Annexin V Conjugates for Apoptosis Detection
Annexin V Conjugates for Apoptosis Detection
Citations & References (9)
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
A35122410／455405Pacific Blue
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
製品番号（カタログ番号） A35122
価格（JPY）
99,100
Each
お問い合わせください ›
励起／発光:
410／455
フローサイトメーターレーザーライン:
405
コンジュゲート:
Pacific Blue
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
色青色
概要フローサイトメトリー用アネキシンV、Pacific Blueコンジュゲート
励起／発光410／455
フローサイトメーターレーザーライン405
使用対象 (装置)フローサイトメーター
キット内容1バイアルのアネキシンV、Pacific Blueコンジュゲートを含みます。
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Pacific Blue
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 (9)

引用および参考文献
Abstract
Hepatitis B virus X protein targets Bcl-2 proteins to increase intracellular calcium, required for virus replication and cell death induction.
Authors:Geng X, Huang C, Qin Y, McCombs JE, Yuan Q, Harry BL, Palmer AE, Xia NS, Xue D,
Journal:Proc Natl Acad Sci U S A
PubMed ID:23091012
'Infection with the hepatitis B virus (HBV) promotes the development of hepatitis, cirrhosis, and hepatocellular carcinoma (HCC) and is a leading cause of morbidity and mortality worldwide. HBV X protein (HBx) is an important effector for HBV pathogenesis, but its cellular targets and acting mechanisms remain elusive. We show here ... More
Live attenuated lentivirus infection elicits polyfunctional simian immunodeficiency virus Gag-specific CD8+ T cells with reduced apoptotic susceptibility in rhesus macaques that control virus replication after challenge with pathogenic SIVmac239.
Authors:Genescà M, Rourke T, Li J, Bost K, Chohan B, McChesney MB, Miller CJ,
Journal:J Immunol
PubMed ID:17878372
'HIV-specific CD8+ T cells that secrete multiple cytokines in response to Ag stimulation are associated with the control of virus replication during chronic HIV infection. To determine whether the presence of polyfunctional CD8+ T cell responses distinguishes protected and unprotected monkeys in a live attenuated lentivirus model, SIV Gag peptide-specific ... More
A novel method for long term bone marrow culture and genetic modification of murine neutrophils via retroviral transduction.
Authors:Zemans RL, Briones N, Young SK, Malcolm KC, Refaeli Y, Downey GP, Worthen GS,
Journal:J Immunol Methods
PubMed ID:19010330
'Neutrophils are a critical component of the innate immune response to invading microbial pathogens. However, an excessive and/or prolonged neutrophil response can result in tissue injury that is thought to underlie the pathogenesis of various inflammatory diseases. The development of novel therapeutic strategies for inflammatory diseases depends on an improved ... More
Modified vaccinia virus Ankara-based vaccine vectors induce apoptosis in dendritic cells draining from the skin via both the extrinsic and intrinsic caspase pathways, preventing efficient antigen presentation.
Authors:Guzman E, Cubillos-Zapata C, Cottingham MG, Gilbert SC, Prentice H, Charleston B, Hope JC,
Journal:J Virol
PubMed ID:22419811
'Dendritic cells (DC) are potent antigen-presenting cells and central to the induction of immune responses following infection or vaccination. The collection of DC migrating from peripheral tissues by cannulation of the afferent lymphatic vessels provides DC which can be used directly ex vivo without extensive in vitro manipulations. We have ... More
The phosphatidylinositol-3-kinase inhibitor NVP-BKM120 overcomes resistance signals derived from microenvironment by regulating the Akt/FoxO3a/Bim axis in chronic lymphocytic leukemia cells.
Authors:Rosich L, Saborit-Villarroya I, López-Guerra M, Xargay-Torrent S, Montraveta A, Aymerich M, Villamor N, Campo E, Pérez-Galán P, Roué G, Colomer D,
Journal:
PubMed ID:23850807
Phosphatidylinositol-3-kinase pathway is constitutively activated in chronic lymphocytic leukemia mainly due to microenvironment signals, including stromal cell interaction and CXCR4 and B-cell receptor activation. Because of the importance of phosphatidylinositol-3-kinase signaling in chronic lymphocytic leukemia, we investigated the activity of the NVP-BKM120, an orally available pan class I phosphatidylinositol-3-kinase inhibitor. ... More
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