Annexin V Conjugates for Apoptosis Detection
Annexin V Conjugates for Apoptosis Detection
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。
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
A13202578/603532、561Alexa Fluor 568
A13201495/519488Alexa Fluor 488
A13199494/518488FITC
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) A13202
価格(JPY)
112,400
Each
お問い合わせください ›
励起/発光:
578/603
フローサイトメーターレーザーライン:
532、561
コンジュゲート:
Alexa Fluor 568
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
オレンジ-赤
概要アネキシンV、Alexa Fluor 568コンジュゲート
励起/発光578/603
フローサイトメーターレーザーライン532、561
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 568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 568
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.

引用および参考文献 (14)

引用および参考文献
Abstract
Quantitation of microparticles released from coated-platelets.
Authors:Dale GL, Remenyi G, Friese P,
Journal:J Thromb Haemost
PubMed ID:16102115
'Dual agonist stimulation of platelets with thrombin and convulxin results in generation of coated-platelets, a sub-population of cells known formerly as COAT-platelets (collagen and thrombin). Coated-platelets retain several procoagulant proteins on their surface and express phosphatidylserine (PS). In this report, we utilize a new methodology to demonstrate that coated-platelets also ... More
Bacterium-generated nitric oxide hijacks host tumor necrosis factor alpha signaling and modulates the host cell cycle in vitro.
Authors:Mocca B, Wang W,
Journal:J Bacteriol
PubMed ID:22636782
'In mammalian cells, nitric oxide (NO·) is an important signal molecule with concentration-dependent and often controversial functions of promoting cell survival and inducing cell death. An inducible nitric oxide synthase (iNOS) in various mammalian cells produces higher levels of NO· from l-arginine upon infections to eliminate pathogens. In this study, ... More
Accumulation of glycosphingolipids in Niemann-Pick C disease disrupts endosomal transport.
Authors:te Vruchte D, Lloyd-Evans E, Veldman RJ, Neville DC, Dwek RA, Platt FM, van Blitterswijk WJ, Sillence DJ
Journal:J Biol Chem
PubMed ID:15078881
'Glycosphingolipids are endocytosed and targeted to the Golgi apparatus but are mistargeted to lysosomes in sphingolipid storage disorders. Substrate reduction therapy utilizes imino sugars to inhibit glucosylceramide synthase and potentially abrogate the effects of storage. Niemann-Pick type C (NPC) disease is a disorder of intracellular transport where glycosphingolipids (GSLs) and ... More
Phosphatidylserine exposure in B lymphocytes: a role for lipid packing.
Authors:Elliott JI, Sardini A, Cooper JC, Alexander DR, Davanture S, Chimini G, Higgins CF,
Journal:Blood
PubMed ID:16684961
'Plasma membrane lipids are usually distributed asymmetrically, with phosphatidylserine (PS) confined to the inner leaflet. PS exposure at the outer leaflet occurs early in apoptosis, but it is also constitutive on some nonapoptotic cell populations where it plays a role in cell signaling. How PS is transported ("flopped") to the ... More
A ribonucleotide reductase homolog of cytomegalovirus and endothelial cell tropism.
Authors:Brune W, Ménard C, Heesemann J, Koszinowski UH
Journal:Science
PubMed ID:11209080
Human cytomegalovirus infects vascular tissues and has been associated with atherogenesis and coronary restenosis. Although established laboratory strains of human cytomegalovirus have lost the ability to grow on vascular endothelial cells, laboratory strains of murine cytomegalovirus retain this ability. With the use of a forward-genetic procedure involving random transposon mutagenesis ... More