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Antibody Labeling Kits |
Brighter insights, precise research
Tell your cell story with Invitrogen antibody labeling kits
- Reactive dyes—for “do-it-yourself” antibody conjugation, choose from a wide variety of colors with the protein labeling reagent selection guide
- Antibody labeling kits—take advantage of preoptimized protocols and kits, including everything needed to enable your successful labeling experiment. Choose the right kit for your experiment using the antibody labeling kit selection tool
- Custom fluorescent antibody labeling—expert antibody conjugation that gives an excellent conjugation outcome
In addition to exceptional brightness and photostability, our customers can get access to an experienced, problem-solving technical support team, application and experimental tips, and protocols to help with experimental planning.
SiteClick Duet and Quartet antibody modification kits
Precise antibody conjugation in as little as four hours.
Not sure which type of conjugation kit to use? Explore the table and sections below to find the right antibody conjugation kit for your application.
Which antibody labeling kit do you need?
これらのキットは、15分以内に準備できる迅速なプロトコルとコンジュゲートを含んでいます。
| Zip Rapid Antibody Labeling Kits | Zenon Antibody Labeling Kits | |
|---|---|---|
| ラベルされるIgG量／反応* | 100 µg | 1～20 µg |
| ラベリングターゲット／方法 | 遊離リジン／共有結合アミン反応性ケミストリー | IgGのFc部分／抗体アフィニティー |
| degree of labelingの測定 | 不可 | 不可 |
| コンジュゲートの24時間以上の保存 | 可 | 不可 |
| 部位特異的ラベリング | 不可 | 不可 |
| サンプルバッファー中のBSAや他の安定剤との適合性 | なし | あり |
| ラベリング後の精製の必要性 | なし | なし |
| プロトコルに必要な時間 | 15分 | 5分 |
| 適したアプリケーション** | FC、IF、WB、HCA | FC、IF、WB、HCA |
| *キットは、特に記載がない限り、分子量150 kDaの全IgGタンパク質用に至適化されています。これらのキットについては、最適な色素：タンパク質比を得るためのタンパク質濃度調製方法に関する説明書をご確認ください。 | ||
| **FC=フローサイトメトリー、IF=免疫蛍光、WB=ウェスタンブロット分析、HCA=ハイコンテント解析 | ||
これらのキットは、抗体上の一級アミンを共有結合でラベルする従来のアミン反応性色素を使用しています。
| Alexa Fluor Antibody Labeling Kits | Zip Rapid Antibody Labeling Kits | Large-scale antibody labeling kits | ReadyLabel Antibody Labeling Kits | |
|---|---|---|---|---|
| Amount IgG labeled/reaction* | 100 µg | 100 µg | 1 mg | 20 or 100 µg |
| Label target/method | Free lysines/covalent amine-reactive chemistry | |||
| Can measure degree of labeling? | Yes | No | Yes | Yes |
| Can store conjugate more than 24 hr? | Yes | Yes | Yes | Yes |
| Site-specific label? | No | No | No | No |
| Compatible with BSA or other stabilizers in sample buffer? | No | No | No | Yes |
| Requires post-label purification? | Yes - included | No | Yes - included | Yes - included |
| Time required for protocol | 30 min | 15 min | 60 min | 60 min |
| Optimal applications** | FC, IF, WB, HCA | FC, IF, WB, HCA | FC, IF, WB, HCA | FC, IF, WB, HCA |
| *Kits are optimized for whole IgG protein, molecular weight 150 kDa unless otherwise indicated. For those kits, please see instructions on how to adjust protein concentration for optimal dye:protein ratios. | ||||
| **FC=flow cytometry; IF=immunofluorescence; WB=western blot analysis; HCA=high content analysis | ||||
これらの方法は抗原結合部位のラベリングを防ぎます。
| SiteClick Antibody Labeling Kits | SureClick Antibody Labeling Kit | |
|---|---|---|
| Amount IgG labeled/reaction* | 100 µg–5 mg | 100 µg–5 mg |
| Label target/method | Fc glycan of IgG antibodies | Fc domain of human or rabbit IgG antibodies |
| Can measure degree of labeling? | Yes | Yes |
| Can store conjugate more than 24 hr? | Yes | Yes |
| Site-specific label? | Yes | Yes |
| Compatible with BSA or other stabilizers in sample buffer? | Yes | Yes |
| Requires post-label purification? | Yes—included | Yes—included |
| Time required for protocol | Duet kits: 6–8 hours; Quartet kits: 16–18 hours | 90 minutes |
| Optimal applications** | FC, IF, WB, HCA | FC, IF, WB, HCA |
| *Kits are optimized for whole IgG protein, molecular weight 150 kDa unless otherwise indicated. For those kits, please see instructions on how to adjust protein concentration for optimal dye:protein ratios. | ||
| **FC=flow cytometry; IF=immunofluorescence; WB=western blot analysis; HCA=high content analysis | ||
治療用抗体開発における迅速なスクリーニングにも有効な抗体ラベリングキット
| Zenon Antibody Labeling Kits | ReadyLabel Antibody Labeling Kits | |
|---|---|---|
| Amount IgG labeled/reaction* | 1–20 µg (96 labelings) | 20 or 100 µg (5 labelings) |
| Label target/method | Fc portion of IgG/antibody affinity | Free lysines/covalent amine-reactive chemistry |
| Can measure degree of labeling? | No | Yes |
| Can store conjugate more than 24 hr? | No | Yes |
| Site-specific label? | No | No |
| Compatible with BSA or other stabilizers in sample buffer? | Yes | Yes |
| Requires post-label purification? | No | Yes—included |
| Time required for protocol | 5 min | 60 min |
| Optimal applications** | FC, IF, WB, HCA | FC, IF, WB, HCA |
| *Kits are optimized for whole IgG protein, molecular weight 150 kDa unless otherwise indicated. For those kits, please see instructions on how to adjust protein concentration for optimal dye:protein ratios. | ||
| **FC=flow cytometry; IF=immunofluorescence; WB=western blot analysis; HCA=high content analysis | ||
pHrodo色素は中性pHでは非蛍光性で、インターナリゼーションされると酸性環境で明るい蛍光を発します。
| SiteClick Lysolight Labeling Kits | Zenon Lysolight Labeling Kits | pHrodo Antibody Labeling Kits | SiteClick pHrodo Antibody Labeling Kits | Zenon pHrodo Antibody Labeling Reagents | |
|---|---|---|---|---|---|
| Amount IgG labeled/reaction* | 100 µg–5 mg | 1–20 µg | 20 µg–1 mg | 100 µg–5 mg | 1–20 µg |
| Label target/method | Fc portion of IgG | Fc portion of IgG/antibody affinity | Free lysines/covalent amine-reactive chemistry | Fc portion of IgG | Fc portion of IgG/antibody affinity |
| Can measure degree of labeling? | Yes | No | Yes | Yes | No |
| Can store conjugate more than 24 hr? | Yes | No | Yes | Yes | No |
| Site-specific label? | Yes | No | No | Yes | No |
| Compatible with BSA or other stabilizers in sample buffer? | Yes | Yes | No | Yes | Yes |
| Requires post-label purification? | Yes | No | Yes—included | Yes—included | No |
| Time required for protocol | 6 hours | 5 min | 60 min | Overnight incubation | 5 min |
| Optimal applications** | FC, IF, WB, HCA | FC, IF, WB, HCA | FC, IF, WB, HCA | FC, IF, WB, HCA | FC, IF, WB, HCA |
| *Kits are optimized for whole IgG protein, molecular weight 150 kDa unless otherwise indicated. For those kits, please see instructions on how to adjust protein concentration for optimal dye:protein ratios. | |||||
| **FC=flow cytometry; IF=immunofluorescence; WB=western blot analysis; HCA=high content analysis | |||||
これらのキットは、15分以内に準備できる迅速なプロトコルとコンジュゲートを含んでいます。
| Zip Rapid Antibody Labeling Kits | Zenon Antibody Labeling Kits | |
|---|---|---|
| ラベルされるIgG量／反応* | 100 µg | 1～20 µg |
| ラベリングターゲット／方法 | 遊離リジン／共有結合アミン反応性ケミストリー | IgGのFc部分／抗体アフィニティー |
| degree of labelingの測定 | 不可 | 不可 |
| コンジュゲートの24時間以上の保存 | 可 | 不可 |
| 部位特異的ラベリング | 不可 | 不可 |
| サンプルバッファー中のBSAや他の安定剤との適合性 | なし | あり |
| ラベリング後の精製の必要性 | なし | なし |
| プロトコルに必要な時間 | 15分 | 5分 |
| 適したアプリケーション** | FC、IF、WB、HCA | FC、IF、WB、HCA |
| *キットは、特に記載がない限り、分子量150 kDaの全IgGタンパク質用に至適化されています。これらのキットについては、最適な色素：タンパク質比を得るためのタンパク質濃度調製方法に関する説明書をご確認ください。 | ||
| **FC=フローサイトメトリー、IF=免疫蛍光、WB=ウェスタンブロット分析、HCA=ハイコンテント解析 | ||
これらのキットは、抗体上の一級アミンを共有結合でラベルする従来のアミン反応性色素を使用しています。
| Alexa Fluor Antibody Labeling Kits | Zip Rapid Antibody Labeling Kits | Large-scale antibody labeling kits | ReadyLabel Antibody Labeling Kits | |
|---|---|---|---|---|
| Amount IgG labeled/reaction* | 100 µg | 100 µg | 1 mg | 20 or 100 µg |
| Label target/method | Free lysines/covalent amine-reactive chemistry | |||
| Can measure degree of labeling? | Yes | No | Yes | Yes |
| Can store conjugate more than 24 hr? | Yes | Yes | Yes | Yes |
| Site-specific label? | No | No | No | No |
| Compatible with BSA or other stabilizers in sample buffer? | No | No | No | Yes |
| Requires post-label purification? | Yes - included | No | Yes - included | Yes - included |
| Time required for protocol | 30 min | 15 min | 60 min | 60 min |
| Optimal applications** | FC, IF, WB, HCA | FC, IF, WB, HCA | FC, IF, WB, HCA | FC, IF, WB, HCA |
| *Kits are optimized for whole IgG protein, molecular weight 150 kDa unless otherwise indicated. For those kits, please see instructions on how to adjust protein concentration for optimal dye:protein ratios. | ||||
| **FC=flow cytometry; IF=immunofluorescence; WB=western blot analysis; HCA=high content analysis | ||||
これらの方法は抗原結合部位のラベリングを防ぎます。
| SiteClick Antibody Labeling Kits | SureClick Antibody Labeling Kit | |
|---|---|---|
| Amount IgG labeled/reaction* | 100 µg–5 mg | 100 µg–5 mg |
| Label target/method | Fc glycan of IgG antibodies | Fc domain of human or rabbit IgG antibodies |
| Can measure degree of labeling? | Yes | Yes |
| Can store conjugate more than 24 hr? | Yes | Yes |
| Site-specific label? | Yes | Yes |
| Compatible with BSA or other stabilizers in sample buffer? | Yes | Yes |
| Requires post-label purification? | Yes—included | Yes—included |
| Time required for protocol | Duet kits: 6–8 hours; Quartet kits: 16–18 hours | 90 minutes |
| Optimal applications** | FC, IF, WB, HCA | FC, IF, WB, HCA |
| *Kits are optimized for whole IgG protein, molecular weight 150 kDa unless otherwise indicated. For those kits, please see instructions on how to adjust protein concentration for optimal dye:protein ratios. | ||
| **FC=flow cytometry; IF=immunofluorescence; WB=western blot analysis; HCA=high content analysis | ||
治療用抗体開発における迅速なスクリーニングにも有効な抗体ラベリングキット
| Zenon Antibody Labeling Kits | ReadyLabel Antibody Labeling Kits | |
|---|---|---|
| Amount IgG labeled/reaction* | 1–20 µg (96 labelings) | 20 or 100 µg (5 labelings) |
| Label target/method | Fc portion of IgG/antibody affinity | Free lysines/covalent amine-reactive chemistry |
| Can measure degree of labeling? | No | Yes |
| Can store conjugate more than 24 hr? | No | Yes |
| Site-specific label? | No | No |
| Compatible with BSA or other stabilizers in sample buffer? | Yes | Yes |
| Requires post-label purification? | No | Yes—included |
| Time required for protocol | 5 min | 60 min |
| Optimal applications** | FC, IF, WB, HCA | FC, IF, WB, HCA |
| *Kits are optimized for whole IgG protein, molecular weight 150 kDa unless otherwise indicated. For those kits, please see instructions on how to adjust protein concentration for optimal dye:protein ratios. | ||
| **FC=flow cytometry; IF=immunofluorescence; WB=western blot analysis; HCA=high content analysis | ||
pHrodo色素は中性pHでは非蛍光性で、インターナリゼーションされると酸性環境で明るい蛍光を発します。
| SiteClick Lysolight Labeling Kits | Zenon Lysolight Labeling Kits | pHrodo Antibody Labeling Kits | SiteClick pHrodo Antibody Labeling Kits | Zenon pHrodo Antibody Labeling Reagents | |
|---|---|---|---|---|---|
| Amount IgG labeled/reaction* | 100 µg–5 mg | 1–20 µg | 20 µg–1 mg | 100 µg–5 mg | 1–20 µg |
| Label target/method | Fc portion of IgG | Fc portion of IgG/antibody affinity | Free lysines/covalent amine-reactive chemistry | Fc portion of IgG | Fc portion of IgG/antibody affinity |
| Can measure degree of labeling? | Yes | No | Yes | Yes | No |
| Can store conjugate more than 24 hr? | Yes | No | Yes | Yes | No |
| Site-specific label? | Yes | No | No | Yes | No |
| Compatible with BSA or other stabilizers in sample buffer? | Yes | Yes | No | Yes | Yes |
| Requires post-label purification? | Yes | No | Yes—included | Yes—included | No |
| Time required for protocol | 6 hours | 5 min | 60 min | Overnight incubation | 5 min |
| Optimal applications** | FC, IF, WB, HCA | FC, IF, WB, HCA | FC, IF, WB, HCA | FC, IF, WB, HCA | FC, IF, WB, HCA |
| *Kits are optimized for whole IgG protein, molecular weight 150 kDa unless otherwise indicated. For those kits, please see instructions on how to adjust protein concentration for optimal dye:protein ratios. | |||||
| **FC=flow cytometry; IF=immunofluorescence; WB=western blot analysis; HCA=high content analysis | |||||
キットをお探しではない方、または別の色素が必要な方
フローサイトメトリー抗体のコンジュゲーションについては、狭い蛍光スペクトル範囲と最小限のクロスレーザー励起によりさらに高い分解能を実現するよう設計されたイムノフェノタイピング用NovaFluor Dyeをご覧ください。
DyLight蛍光色素を使用して1 mgまたは100 μgの抗体をラベリングするためにはDyLight抗体ラベリングキットをご確認ください。
スタンドアロンのアミン反応性色素やアミン反応性ケミストリに関する詳細は、蛍光色素とアミン反応性誘導体—第1章、Molecular Probesハンドブックをご覧ください。
スタンドアロン試薬としてのチオール反応性色素も提供しています。これらの詳細についてはチオール反応性プローブ—第2章、Molecular Probesハンドブックを参照してください。
Amine-reactive antibody labeling kits
Zip Rapid Antibody Labeling Kits
Zip Alexa Fluor Rapid Antibody Labeling Kitを使用すると、貴重な抗体をAlexa Flour色素で効率的にラベリングして、15分以内に使用できる抗体コンジュゲートを作成できます。
- 100 μgのIgG抗体をラベリング
- キットあたり3回分のラベリング反応
- 非常に迅速で使用が簡単
- 100%の抗体回収率—精製ステップは必要なし
- 化学やコンジュゲーションの経験は必要なし
このキットには、3回分のラベリング反応を行うために必要なすべてのものが含まれています。共有結合でラベルされたコンジュゲートは、フローサイトメトリー、蛍光顕微鏡、免疫組織化学、一次抗体検出、ELISA、免疫細胞化学、間接FISHなどの複数のアプリケーションに適しています。Zipバッファーを含有するコンポーネントAに1 mLの水を加え、さらに1 mgの抗体を加えて、15分間インキュベートするだけで、抗体をすぐに使用できます（図1）。
図をクリックして拡大
図1.Zip Rapid Antibody Labeling Kitの概要
Zip Rapid Antibody Labeling Kit選択ガイド
励起／蛍光=最大励起および蛍光（nm）
Alexa Fluor色素およびその他の蛍光色素の詳細は蛍光色素選択ガイドご確認ください
Alexa Fluor Antibody Labeling Kits
Invitrogen Alexa Fluor Antibody labeling kits are easy-to-use kits to covalently label primary antibodies with Alexa Fluor dyes. Each antibody labeling kit includes everything needed to perform up to three separate antibody conjugation reactions. The kits also now include pre-assembled and pre-filled spin columns to more easily and quickly purify out free dye from your sample.
- Label 100 µg of antibody per reaction
- Three labeling reactions per kit
- Labeled antibodies ready in ~30 minutes
- ~15 minutes actual hands-on time
- Helps save time and minimize error with pre-assembled and pre-filled post-label purification columns
- Requires removal of BSA and other protein stabilizers before conjugation
Alexa Fluor Antibody Labeling kits use amine-reactive chemistry to covalently attach Alexa Fluor dyes or a biotin molecule to your IgG antibody. For best results, ensure your primary antibody is in a compatible buffer, free of BSA or other protein stabilizers. If your antibody of interest needs clean-up, you can achieve this using antibody clean-up kits (Cat. No 89950, Cat No. 78609, or Cat No. 45206)
The first step in the conjugation workflow (Figure 2) is adding the included sodium bicarbonate to the primary antibody solution. This prepares the antibody for conjugation by enhancing the reactivity of the antibody's lysine residues.
The second step is to mix this solution with the included amine-reactive Alexa Fluor dye or biotin compound. The reactive groups on the fluorescent dye will attach to the primary amines on the IgG antibody's side chains. This creates a stable amide bond that covalently conjugates the fluorescent dye or biotin to the antibody. The conjugation reaction completes efficiently at room temperature after 15 minutes of incubation.
The final step is to remove the free or unbound dye which helps reduce background signal in your downstream applications. Each kit includes improved purification spin columns to help you achieve this. The pre-filled, pre-assembled columns are now compatible with standard microcentrifuges, helping save you time and lab space.
Click image to enlarge
Figure 2. Overview of Alexa Fluor Antibody Labeling Kits process. Simply adjust the protein concentration to ~1 mg/mL in the provided buffer, then add it to a vial of amine-reactive dye. Purification is accomplished on the included Zeba Dye and Biotin removal columns optimized for ≥7 kDa proteins.
Antibody labeling kits selection guide
| Label | Ex/Em | Cat. No. |
|---|---|---|
| Alexa Fluor 350 | 346/442 | A88061 |
| Alexa Fluor 488 | 494/519 | A88062 |
| Alexa Fluor 532 | 530/554 | A88063 |
| Alexa Fluor 546 | 554/570 | A88064 |
| Alexa Fluor 555 | 555/565 | A88065 |
| Alexa Fluor 568 | 577/603 | A88066 |
| Alexa Fluor 594 | 590/617 | A88067 |
| Alexa Fluor 647 | 650/668 | A88068 |
| Alexa Fluor 680 | 679/702 | A88069 |
| Alexa Fluor 790 | 785/810 | A88070 |
| Biotin | N/A | B88060 |
| Pacific Blue dye | 410/455 | P30013 |
| Pacific Orange Dye | 405/551 | P30014 |
励起／蛍光=最大励起および蛍光（nm）
Alexa Fluor色素およびその他の蛍光色素の詳細は蛍光色素選択ガイドご確認ください
ReadyLabel Antibody Labeling Kits
Invitrogen ReadyLabel Antibody Labeling Kits provide a 2-in-1 antibody labeling and purification kit, to label antibodies directly in presence of impurities like BSA and other protein stabilizers without compromising signal to background.
Our ReadyLabel dyes are now available in expanded dye formats optimized for spectral unmixing, to enable primary antibody detection for your multiplex or spatial biology experiments.
- No antibody purification step needed
- Label 20 μg or 100 μg of IgG antibody
- 5 labeling reactions per kit
- Labeled antibodies ready in 60 minutes
- Single or multiplex staining
- Improved signal to background
ReadyLabel Antibody Labeling Kits allow for fast, convenient antibody conjugation of off-the-shelf antibodies and cell culture supernatants without prior purification of the antibody solution. This works because the provided ReadyLabel spin columns contain resin that filter out contaminating stabilizer proteins present in the antibody solution (Figure 3). The resulting covalently bonded antibody conjugate can be used with flow cytometry, western blotting, ELISA, and indirect FISH.
ReadyLabel kits are equipped with your choice of our most popular stains to produce high yields of bright, soluble conjugates. Need a different dye? Consider our ReadyLabel Flex Kits, which contain everything needed except the dye, and use a dye of your choice. Our most popular ReadyLabel Kits (Alexa Fluor 488, Alexa Fluor 647, Biotin, and Flex) are available in two different pack sizes (20 μg or 100 μg IgG per reaction) and include all components necessary for five labeling reactions. Our ReadyLabel kits designed for high-plex staining or spatial biology workflows are available in the 20 μg pack size.
Figure 3. Antibodies are often stored with BSA or other protein stabilizers to prevent degradation. ReadyLabel kits are equipped with resin-containing spin columns that capture these contaminants and allow you to skip the purification step of your antibody solution. With approximately 20 minutes of hands-on time, and 30 minutes of incubation at room temperature, your covalently bonded antibody conjugate is now ready to use.
How to: ReadyLabel Antibody Labeling Kits protocol
Ordering information for ReadyLabel Antibody Labeling Kits
ReadyLabel Antibody Labeling Kits offer an exceptional advantage for multiplex or high-plex experiments. Unlike secondary detection methods, which are often constrained by the number of available IgG species, our kits allow you to directly conjugate your primary antibodies of your choice, filling gaps in your panels. This means you can achieve higher levels of multiplexing with greater flexibility.
Designed with high-quality, bright dyes, optimized for spectral unmixing, and a focus on minimizing background noise through BSA removal, our kits enable clear and precise signals in your experiments. This clarity enhances the reliability and accuracy of your results, making ReadyLabel Antibody Labeling Kits well suited for multiplexing applications.
Visit our spatial biology resource center to learn more
Download the Application Note: Streamlined spatial analysis of the brain tumor microenvironment and associated pathological states using multiplex immunohistochemistry
Figure 4. Imaged on the EVOS S1000 Spatial Imaging Instrument at 20x, FFPE human colon tissue was stained using ReadyLabel Kits. ReadyLabel Alexa Fluor 420 SMA, ReadyLabel Alexa Fluor 514 CD20, ReadyLabel Alexa Fluor Plus 555 PCNA, ReadyLabel Alexa Fluor 594 CD45RB, ReadyLabel Alexa Fluor 647 eCadherin, ReadyLabel Alexa Fluor 700 Cytokeratin, and ReadyLabel Alexa Fluor Plus 750 CD68.
Figure 5. Imaged on the EVOS S1000 Spatial Imaging Instrument at 20x, FFPE Human brain tissue with Parkinson’s disease was stained using a mix of conjugated primary antibodies and ReadyLabel kits. DAPI, Alexa Fluor 420 CD68, Alexa Fluor 488 GFAP, Alexa Fluor Plus 555 Ki67, ReadyLabel Alexa Fluor 594 LRRK2, ReadyLabel Alexa Fluor 647 alpha Synuclein, Alexa Fluor 700 p53, and Alexa Fluor Plus 750 Vimentin.
Figure 6. Imaged on the EVOS S1000 Spatial Imaging Instrument at 20x, FFPE human tonsil tissue was stained using ReadyLabel Kits. ReadyLabel Alexa Fluor 420 SMA, ReadyLabel Alexa Fluor 514 CD20, ReadyLabel Alexa Fluor Plus 555 PCNA, ReadyLabel Alexa Fluor 594 CD45RB, ReadyLabel Alexa Fluor 647 eCadherin, ReadyLabel Alexa Fluor 700 Cytokeratin, and ReadyLabel Alexa Fluor Plus 750 CD68.
Figure 7. Imaged on the EVOS S1000 Spatial Imaging Instrument at 20x, FFPE human colon tissue was stained using ReadyLabel Kits. ReadyLabel Alexa Fluor 420 GranzymeB, ReadyLabel Alexa Fluor 514 Cytokeratin, ReadyLabel Alexa Fluor Plus 555 PCNA, ReadyLabel Alexa Fluor 594 HLA-DR, ReadyLabel Alexa Fluor 647 eCadherin, ReadyLabel Alexa Fluor 700 SMA, and ReadyLabel Alexa Fluor Plus 750 CD68.
Large-scale antibody labeling kits
Thermo Fisher Scientific large-scale antibody or protein labeling kits provide a nearly effortless way to covalently label 1 mg of IgG antibody with a fluorescent dye (including Alexa Fluor dyes) or biotin.
- Label 1 mg antibody
- Three labeling reactions in each kit
- Removal of BSA and other stabilizers is required prior to labeling
- Not compatible with amine-containing buffers (e.g., Tris)
- Ready to use in 2 hours
- ~ 30 minutes actual hands-on time
The large-scale antibody or protein labeling kits utilize an amine-reactive fluorophore or hapten to covalently attach the label to the IgG antibody. Once formed, the covalent bond between the label and the protein is extremely stable—you are using the same chemistry we use to prepare our own primary and secondary conjugates.
Simply add ~1 mg of purified IgG (in ~500 μL and free of amine-containing buffers such as Tris) to one of the vials containing a premeasured quantity of amine-reactive dye and a magnetic stir bar. Purification is accomplished by a pre-packed spin column supplied with the kit and affords high antibody recovery (typically >85%). The entire labeling and purification procedure can be completed in as little as 90 minutes (approximately 15 mins hands-on time). Removal of free dye is essential for the determination of the degree of labeling (DOL). Everything needed to perform three conjugations is provided.
図をクリックして拡大
Figure 5. Overview of the large-scale antibody labeling kits process. Adjust the protein concentration to ~2 mg/mL, add provided bicarbonate, then add the solution to the vial of reactive dye. Purification is accomplished using a pre-packed spin column that is compatible with proteins ≥20 kDa.
Large-scale antibody or protein labeling kits selection guide
| 標識 | 励起／蛍光 | カタログ番号 |
|---|---|---|
| Alexa Fluor 350 | 346／442 | A10170 |
| Alexa Fluor 488 | 495／519 | A10235 |
| Alexa Fluor 532 | 532／553 | A10236 |
| Alexa Fluor 546 | 556／573 | A10237 |
| Alexa Fluor 555 | 555／565 | A20174 |
| Alexa Fluor 568 | 578／603 | A10238 |
| Alexa Fluor 594 | 590／617 | A10239 |
| Alexa Fluor 633 | 621／639 | A20170 |
| Alexa Fluor 647 | 650／665 | A20173 |
| Alexa Fluor 660 | 663／690 | A20171 |
| Alexa Fluor 680 | 679／702 | A20172 |
| Fluorescein | 494／518 | F10240 |
| Oregon Green 488 | 496／524 | O10241 |
| Pacific Blue色素 | 410／455 | P30012 |
| Pacific Orange色素 | 400／551 | P30016 |
| Texas Red色素 | 595／615 | T10244 |
| ビオチン | NA | D20655 |
励起／蛍光=最大励起および蛍光（nm）
Alexa Fluor色素およびその他の蛍光色素の詳細は蛍光色素選択ガイドご確認ください
Affinity antibody labeling kits
Zenon Antibody Labeling Kits
Invitrogen Zenonラベリング技術は、ハイブリドーマ培養上清のような少量の未精製ソースからでも、抗体をさまざまな蛍光色素に迅速に結合させます。Zenonフラグメントは一次抗体のFc部位に結合するように設計されており、抗体の機能を損なうことなく、少量の抗体を特異的に非共有結合で標識することができます。Zenon Alexa Fluor Plus labeling reagentsは、ZenonテクノロジーのスピードとスケーラビリティをAlexa Fluor Plus色素の優れた性能と融合させ、抗体医薬探索における抗体結合のスクリーニングに最適なシステムを提供します。
- インキュベーション時間は5分
- ラベリング反応当たりの一次抗体は1～20 µg
- BSAやその他の安定化タンパク質に適合
- 精製ステップは不要
- 抗体結合の迅速なスクリーニングを実現
- 一次抗体の機能を維持
Invitrogen Zenon labeling reagentsは、ヒト、マウス、およびウサギIgG抗体をさまざまなフルオロフォアで非共有的にラベルする、迅速で使いやすいシステムを提供します。ラベル済みZenon FabフラグメントはIgG抗体のFc部位に迅速に結合するように設計されており、抗体の機能を維持します。Zenon Fab-抗体結合は5分未満で起こり、必要な出発物質は少量で、精製ステップは不要です。
図6.マウスまたはヒトIgGをZenon標識試薬で室温で5分間標識細胞をラベル済みIgGでわずか30分間処理すると、表面抗原を検出でき、最大24時間処理すると、抗体の内在化を確認できます。イメージングと定量は、フローサイトメトリー、ハイコンテントイメージング、または生細胞インキュベーターにより検出および分析できます。
Zenon labeling reagentsを使用した抗体結合の迅速スクリーニング
The Invitrogen Zenon Alexa Fluor Plus labeling reagents have been optimized to screen for antibody binding, giving a bright fluorescence-based indication of antibody binding. These reagents combine the rapid and scalable Zenon technology with the superior Alexa Fluor Plus fluorophores to provide a higher signal to noise ratio and to increase confidence in your screening results.
Want to learn more about screening for binding of therapeutic antibodies?
View our Scientific Poster for Antibody Labeling Reagents for Therapeutic Antibody Screening
図7.さまざまなマウスおよびヒトIgGをZenon Goat Anti-Human Alexa Fluor Plus Labeling Reagentで処理しました。5分間のインキュベーション後、SKBR3細胞を抗体で60分間処理し、CellInsight CX7 LED Proで結合をスクリーニングしました。トラスツズマブは、このスクリーニングの中で、SKBR3細胞の細胞表面に結合することが知られている唯一の抗体でした。
Zenon Antibody Labeling Kits for antibody binding
抗体インターナリゼーションのスクリーニングにご興味がありますか？
Zenon pHrodo Labeling Reagentsをご覧ください
Zenon pHrodo Labeling Reagentsをご覧ください
Zenon Antibody Labeling Kits for multiplex immunofluorescence
| 反応性 | 標的アイソタイプ | 製品リスト |
|---|---|---|
| マウス | IgG1 | すべてのキットを見る |
| ヒト | IgG | すべてのキットを見る |
| ウサギ | IgG | すべてのキットを見る |
Site-specific antibody labeling kits
SiteClick Antibody Labeling Kitsのクリックケミストリーラベリング
Thermo Fisher Scientific SiteClick labeling kits allow simple and gentle site-selective attachment of detection molecules to heavy chain N-linked glycans—far from the antigen-binding domain—providing excellent reproducibility from labeling to labeling and from antibody to antibody. A number of different detection molecules can be site-selectively attached to the heavy chain glycans—including phycobiliproteins (e.g., R-PE), QDot probes, Alexa Fluor dyes, pHrodo dye, metal-chelating compounds, and other small molecules like biotin—allowing multiplex analysis with antibodies from the same species. Features include:
- Labeling of 100 μg up to 5 mg of antibody
- Easy to follow step-by-step protocol
- Highly efficient, site-specific, reproducible labeling chemistry
- Kits and dyes available in a flexible format for easy design of multi-color experiments
一般的にIgG抗体は、抗体の重鎖Fcドメインに位置する特定の保存されたアスパラギン残基に結合した2つのN結合型糖鎖を有しています。これらの糖鎖は主に、2つの末端分岐を持つ複雑な二分岐グリカンであり、構造的に非常に均質で、グリカン分岐の末端配列には非常に高い一貫性があります。抗体のグリカン分岐の大部分は、ガラクトース-N-アセチルグルコサミン（Gal-GlcNAc-）またはN-アセチルグルコサミン（GlcNAc-）で終端しています。β-ガラクトシダーゼにより末端Gal残基を除去することで、大部分の末端GlcNAc標識部位が露出し、その後の酵素的β-ガラクトシルトランスフェラーゼ（GalT）反応を可能にします（図8）。β-ガラクトシダーゼによる末端Gal残基の分解の結果、各N結合型グリカンは重鎖当たり平均して2つの末端GlcNAc残基（抗体当たり4つの末端GlcNAc）を含有します。アジド活性化抗体は、銅フリーのクリックケミストリーを利用して、ジベンゾシクロオクチン（DIBO）官能化プローブでラベリングできます（図9）。
図をクリックして拡大
図8.SiteClick Antibody Labeling Kitsの概要。SiteClick抗体ラベリングプロセスの最初のステップは、β-ガラクトシダーゼを使用して抗体の重鎖N結合グリカンから末端ガラクトース残基を除去し、基本的に修飾可能なすべてのGlcNAc残基を露出させることです。次に、GalT（Y289L）酵素を使用してGalNAzを末端GlcNAc残基に酵素的に結合させることにより、遊離末端GlcNAc残基をアジドタグで活性化します。3番目のステップでは、選択したジベンゾシクロオクチン（DIBO）官能化プローブ（Alexa Fluor 488 sDIBOアルキンなど）とアジド残基を反応させます（この反応については図10を参照）。平均的なラベリング程度は抗体あたり3～3.5標識です。
図9.銅不要クリックケミストリーアジド／DIBO反応。アジド部分とアルキン部分は相互に交換可能です。分子をDIBOでラベリングし、フルオロフォアまたはハプテン-アジドと反応させることが可能です。
SiteClick法は、ヒト、ウサギ、マウス、ラット、ヤギ、ハムスター、ニワトリなど、さまざまな種の抗体に適合します。さらにSiteClickラベリングは、IgG、IgM、およびIgYなどの複数の抗体クラスで有効です；ニワトリIgY抗体は2つではなく6つの重鎖グリカンを有するため、より程度が高いラベリングが可能です。
SiteClick抗体ラベリングシステムは柔軟性を提供します
SiteClick抗体ラベリングシステムには2つのフォーマットがあります：
- 抗体調製用のアジド修飾成分と、抗体をラベリングするためのsDIBOアルキンの両方を含むキット（完全な反応は図8AおよびBに図示）。
- アジド修飾成分のみを含むキット（図8Aに示された反応の最初の2ステップ）。
- 修飾抗体のラベリングに使用できるさまざまなスタンドアロンsDIBOアルキン試薬（図8Bのステップ）。
このようなフォーマットの柔軟性により、同じ抗体のラベリング程度に影響を与えることなく、使用する蛍光色素を容易に交換できることや、必要に応じて修飾抗体の大量のバッチを調製したり少量をラベリングできることなど、いくつかの利点があり、それでいてラベリングや機能の程度の一貫性を維持します。
抗体の機能性は高度に維持されます
The covalent and site-specific attachment of the dyes to the N-linked glycans on the antibody heavy chain Fc domain helps ensure the immunogenicity of the antigen binding site is preserved, resulting in more efficient binding to the antigen as compared to antibodies randomly labeled with amine-reactive dyes. In a side-by-side comparison of SiteClick-labeled and amine-reactive labeled anti-troponin 1 antibody, the sensitivity of the SiteClick-labeled antibody was greater than that of the amine-reactive labeled antibody (Figure 10).
図10.ビーズベースのサンドイッチアッセイにおけるSiteClick抗体ラベリングとアミン反応性抗体ラベリングの比較。抗トロポニン（Trpl）抗体捕捉ビーズを使用して、SiteClick系またはアミン反応性系でラベリングしたビオチン化抗Trpl抗体でトロポニン抗原を検出しました。ストレプトアビジンを介した化学発光アッセイを用いて結合を検出しました。
抗体のラベリング効率におけるロット間の一貫性
Labeling antibodies with the SiteClick antibody labeling system helps give you confidence that the antibody will be labeled the same way every time, with no time needed for reaction optimization. This means the same number of dyes attached to each molecule, the same preservation of the antigen binding site, helping ensure the same results every time. An example of the consistency in labeling between different antibodies is shown in Figure 11.
図11.ロット間および抗体間の一貫性3年間にわたり、β-ガラクトシダーゼで切断されたアジド活性化抗体とSiteClick Alexa Fluor 488 sDIBO Alkyne色素を使用して、まったく同じ条件下で、異なる種の28種類の一次抗体を異なるユーザーによりラベリングしました。degree of labelingは平均して3.4±0.39で、SiteClickラベリングシステムの一貫性を示しています。これらの抗体の種類、種、および抗原性については表1を参照してください。
Table 1. Antigenicity of primary antibodies labeled in degree of labeling study (Figure 11).
| Human IgG1 | Human IgG3 | Mouse IgG1 | Mouse IgG2a | Goat Polyclonal |
|---|---|---|---|---|
| EGFR | Lymphoma cell | GD2 | CD4 | Apolipoprotein-A2 |
| HER2/neu | GD3 | β-tubulin | CD3 | |
| sLea | sLea | CD8a | CD8 | |
| GD2 | f-GM1 | CD45 | CD56 | |
| GM2 | Tn | C3b, iC3b | C3b, iC3b, C3dg | |
| J591 | Interferon-γ |
SiteClick Antibody Labeling Kits選択ガイド
| Product name | Antibody quantity labeled | Ex/Em* | Cat. No. |
|---|---|---|---|
| Kits for antibody modification only | |||
| SiteClick Quartet Antibody Modification Kit | 100 μg | N/A | S11003 |
| 1 mg | S11002 | ||
| 5 mg | S11001 | ||
| 3 x 100 μg | S11007 | ||
| SiteClick Duet Antibody Modification Kit | 100 μg | N/A | S11006 |
| 1 mg | S11005 | ||
| 5 mg | S11004 | ||
| 3 x 100 μg | S11008 | ||
| Standalone labeling reagents | |||
| SiteClick LysoLight Green sDBCO | 100 μg | 488/525 | L36009 |
| 500 μg | L36010 | ||
| LysoLight Deep Red sDBCO | 100 μg | 650/668 | L36011 |
| 500 μg | L36012 | ||
| Zenon LysoLight Green Human IgG Labeling Reagent | 240 µL | 488/525 | Z25627 |
| 1.2 mL | Z25628 | ||
| Zenon LysoLight Green Human IgG Labeling Reagent | 240 µL | 488/525 | Z25629 |
| 1.2 mL | Z25630 | ||
| SiteClick Alexa Fluor 488 sDIBO Alkyne | 100 μg | 495/519 | C20027 |
| 1 mg | S10904 | ||
| 5 mg | S10909 | ||
| SiteClick Alexa Fluor 555 sDIBO Alkyne | 100 μg | 555/565 | C20028 |
| SiteClick Alexa Fluor 647 sDIBO Alkyne | 100 μg | 650/665 | C20029 |
| 1 mg | S10906 | ||
| 5 mg | S10911 | ||
| SiteClick iFL pHrodo Red sDIBO Alkyne | 100 μg | 560/585 | C20034 |
| 1 mg | S10903 | ||
| 5 mg | S10908 | ||
| SiteClick iFL pHrodo Deep Red sDIBO Alkyne | 100 μg | 640/655 | S10914 |
| 1 mg | S10915 | ||
| SiteClick Biotin sDIBO Alkyne | 100 μg | N/A | C20030 |
| 1 mg | S10902 | ||
| 5 mg | S10907 | ||
| SiteClick Amine sDIBO Alkyne | 100 μg | N/A | C20031 |
| Kits containing antibody modification + labeling reagent | |||
| SiteClick R-PE Antibody Labeling kit | 100 μg | 565/578 | S10467 |
| SiteClick Qdot 525 Antibody Labeling kit | 100 μg | 405/525 | S10449 |
| SiteClick Qdot 565 Antibody Labeling kit | 100 μg | 405/565 | S10450 |
| SiteClick Qdot 585 Antibody Labeling kit | 100 μg | 405/585 | S10451 |
| SiteClick Qdot 605 Antibody Labeling kit | 100 μg | 405/605 | S10469 |
| SiteClick Qdot 625 Antibody Labeling kit | 100 μg | 405/625 | S10452 |
| SiteClick Qdot 655 Antibody Labeling kit | 100 μg | 405/655 | S10453 |
| SiteClick Qdot 705 Antibody Labeling kit | 100 μg | 405/705 | S10454 |
| SiteClick Qdot 800 Antibody Labeling kit | 100 μg | 405/800 | S10455 |
| SiteClick Biotin Antibody Labeling Kit | 100 μg | N/A | S20033 |
*励起／蛍光=最大励起および蛍光（nm）
Alexa Fluor色素およびその他の蛍光色素の詳細は蛍光色素選択ガイドご確認ください
リソース