Annexin V Conjugates for Apoptosis Detection
Annexin V Conjugates for Apoptosis Detection
Citations & References (41)
Invitrogen™

Annexin V Conjugates for Apoptosis Detection

アネキシンVスタンドアロンAlexa Fluor、APC、Pacific Blue、PE、FITC、およびフローサイトメトリーを使用したビオチンコンジュゲートでアポトーシスの初期段階を検出します。
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製品番号（カタログ番号）励起／発光フローサイトメーターレーザーラインコンジュゲート
A13201495／519488Alexa Fluor 488
A13199494／518488FITC
A13202578／603532、561Alexa Fluor 568
A13203590／617532Alexa Fluor 594
A13204ビオチン-X
A23202346／442UVAlexa Fluor 350
A23204650／665633～637Alexa Fluor 647
A35108555／565532、561Alexa Fluor 555
A35109679／702633～637Alexa Fluor 680
A35110650/660633～637APC（アロフィコシアニン）
A35111565/578488、532、561PE
A35122410／455405Pacific Blue
製品番号（カタログ番号） A13201
価格（JPY）
112,700
Each
お問い合わせください ›
励起／発光:
495／519
フローサイトメーターレーザーライン:
488
コンジュゲート:
Alexa Fluor 488
アポトーシス検出用のアネキシンVスタンドアロンコンジュゲートにより、初期の細胞アポトーシスを迅速かつ信頼性の高い方法で検出できます。アネキシンVコンジュゲートは、フローサイトメトリーを用いたアポトーシス細胞と非アポトーシス細胞の蛍光シグナル強度の差を最大100倍にします。
アネキシンVはホスファチジルセリン（PS）に対する高い親和性を有しており、アポトーシスを受けている細胞の外側のリーフレットに曝露されます。この親和性により、蛍光標識アネキシンV試薬はアポトーシス研究で一般的に使用されます。

アネキシンVコンジュゲートは、アポトーシスの中間期指標であるホスファチジルセリンの外在化を研究するための迅速で信頼性の高い検出法を提供します。フローサイトメトリーで測定した当社の蛍光アネキシンVコンジュゲートで染色したアポトーシス細胞と非アポトーシス細胞の蛍光強度の差は、通常約100倍です。

Nexins Research社との提携により、当社はAlexa Fluor 350、488、555、568、594、647および680アネキシンVコンジュゲート、アネキシンV APC、Biotin-X、FITC、Pacific Blue、およびPEコンジュゲートなどの最高レベルのアネキシンVコンジュゲート提供します。蛍光性の高いアネキシンVコンジュゲートは、アポトーシスの初期指標の1つであるホスファチジルセリンの外在化を研究するための迅速で信頼性の高い検出法を提供します。

アネキシンV Pacific Blueコンジュゲートはバイオレット励起性であり、バイオレットレーザーを備えた装置や、緑色または赤色蛍光色素を含むマルチカラー実験に最適です。

当社のアネキシンVコンジュゲートの利点：
•より明るいシグナルを得るためのInvitrogen Alexa FluorおよびeFluor色素に対するコンジュゲート
•利用可能なすべてのレーザー用のコンジュゲート
•独立した試薬または使いやすいキットとして利用可能

アポトーシス細胞を検出するためのアネキシンV染色は、生細胞および組織でのみ可能です。サンプルを染色後に固定する場合は、シグナルの一時的な保持を実現するために特定の条件が必要です。これには、アルコールを含まないアルデヒドベースの固定法の使用、Ca2+を含むバッファーの使用、界面活性剤／界面活性剤（洗剤）の回避などが挙げられます。また、アポトーシスアッセイにおけるホスファチジルセリンとアネキシンVとの結合を促進する濃縮アネキシン結合バッファーもご用意しています。

For Research Use Only. Not for use in diagnostic procedures.
仕様
色Green
概要アネキシンV、Alexa Fluor 488コンジュゲート（カタログ番号PHN1010およびPHN1008の代替品）
励起／発光495／519
フローサイトメーターレーザーライン488
使用対象 (装置)フローサイトメーター
キット内容アネキシンV、Alexa Fluor 488コンジュゲートのバイアル1本を含みます。
反応数100
製品タイプアネキシンVコンジュゲート
数量500 μL
出荷条件湿氷
コンジュゲートAlexa Fluor 488
Unit SizeEach
組成および保存条件
冷蔵庫（2℃～8℃）に保存し、遮光してください。

よくあるご質問（FAQ）

I want to study apoptosis using an Annexin V conjugate, but with adherent cells via microscopy instead of flow cytometry. Can this be done?

It has been done, but we don‘t recommend it. Both healthy cells and apoptotic cells possess phosphatidylserine on the cell surface, which can be detected with Annexin V, but apoptotic cells have significantly more of it. You can easily tell the difference between these two populations with flow cytometry, because flow cytometers are more sensitive and have a higher throughput. But with a microscope, you cannot always tell the difference, especially for adherent cells. Instead, for microscopy, we recommend a different technique, such as detecting caspases with CellEvent Caspase Detection Reagents.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I trypsinized my adherent cells and labeled with annexin V, and now my flow data is showing a high percentage of apoptotic cells even for control, untreated cells. What is the problem?

Trypsinization or mechanical scraping of cells temporarily disrupts the plasma membrane, allowing annexin V to bind phosphatidylserine on the cytoplasmic surface of the cell membrane and thus leading to false positive staining. Allow the cells to recover for about 30 minutes in optimal cell culture conditions and medium after trypsinizing/scraping so that they can recover their membrane integrity before staining. For lightly adherent cell lines, such as HeLa and NIH 3T3, another option is to use non-enzyme treatments like Gibco Cell Dissociation Buffer (Cat. No. 13151014).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I detect annexin V staining in an imaging assay?

Annexin V staining is not typically used in imaging experiments; it is a better reagent for flow cytometry analysis. All cells will stain to some extent, so it can be difficult to distinguish a relatively bright annexin V-stained cell from a dimmer non-apoptotic cell. Caspase activation, detected using our CellEvent Caspase 3/7 or Image-iT LIVE Caspase detection kits, is a better method for detecting apoptosis in an imaging assay.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

When should I stain adherent cells with annexin V for flow cytometric analysis? Before or after I trypsinize them?

Trypsinize first and then allow the cells to recover about 30 minutes in optimal cell culture conditions and medium before staining with annexin V conjugates. Trypsinization or mechanical scraping of cells temporarily disrupts the plasma membrane, allowing for annexin V to bind phosphatidylserine on the cytoplasmic surface of the cell membrane and thus leading to false positive staining. For lightly adherent cell lines such as HeLa and NIH 3T3, you could use a less harsh (non-enzymatic) dissociation product like Gibco Cell Dissociation Buffer (Cat. No. 13151014).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I fix my cells after annexin V labeling?

Annexin V staining is best analyzed on live cells. If you need to fix your cells for analysis, then fix in 3.7% formaldehyde in PBS containing calcium and magnesium to maintain binding during fixation. The signal will not be retained after permeabilization, thus annexin V staining is not compatible with internal antibody labeling.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

View all Annexin V Conjugates for Apoptosis Detection FAQs

引用および参考文献 (41)

引用および参考文献
Abstract
VDAC-dependent permeabilization of the outer mitochondrial membrane by superoxide induces rapid and massive cytochrome c release.
Authors:Madesh M, Hajnóczky G
Journal:J Cell Biol
PubMed ID:11739410
'Enhanced formation of reactive oxygen species (ROS), superoxide (O2*-), and hydrogen peroxide (H2O2) may result in either apoptosis or other forms of cell death. Here, we studied the mechanisms underlying activation of the apoptotic machinery by ROS. Exposure of permeabilized HepG2 cells to O2*- elicited rapid and massive cytochrome c ... More
Cyclopentenone prostaglandins of the J series inhibit the ubiquitin isopeptidase activity of the proteasome pathway.
Authors:Mullally JE, Moos PJ, Edes K, Fitzpatrick FA
Journal:J Biol Chem
PubMed ID:11390388
'Electrophilic eicosanoids of the J series, with their distinctive cross-conjugated alpha,beta-unsaturated ketone, inactivate genetically wild type tumor suppressor p53 in a manner analogous to prostaglandins of the A series. Like the prostaglandins of the A series, prostaglandins of the J series have a structural determinant (endocyclic cyclopentenone) that confers the ... More
Aberrant T cell differentiation in the absence of Dicer.
Authors:Muljo SA, Ansel KM, Kanellopoulou C, Livingston DM, Rao A, Rajewsky K
Journal:J Exp Med
PubMed ID:16009718
'Dicer is an RNaseIII-like enzyme that is required for generating short interfering RNAs and microRNAs. The latter have been implicated in regulating cell fate determination in invertebrates and vertebrates. To test the requirement for Dicer in cell-lineage decisions in a mammalian organism, we have generated a conditional allele of dicer-1 ... More
Free cholesterol loading of macrophages induces apoptosis involving the fas pathway.
Authors:Yao PM, Tabas I
Journal:J Biol Chem
PubMed ID:10791964
'Macrophage death is an important feature of atherosclerosis, but the cellular mechanism for this process is largely unknown. There is increasing interest in cellular free cholesterol (FC) excess as an inducer of lesional macrophage death because macrophages accumulate large amounts of FC in vivo, and FC loading of macrophages in ... More
Proteolytic activation of protein kinase C-epsilon by caspase-mediated processing and transduction of antiapoptotic signals.
Authors:Basu A, Lu D, Sun B, Moor AN, Akkaraju GR, Huang J,
Journal:J Biol Chem
PubMed ID:12198125
'Several novel protein kinase C (PKC) isozymes have been identified as substrates for caspase-3. We have previously shown that novel PKCepsilon is cleaved during apoptosis in MCF-7 cells that lack any functional caspase-3. In the present study, we show that in vitro-translated PKCepsilon is processed by human recombinant caspase-3, -7, ... More
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