Mouse thymocytes were surface stained with Anti-Mouse CD4 PE-eFluor® 610 (Product # 61-0042) and Anti-Mouse CD8a PerCP-Cyanine5-5 (Product # 45-0081) followed by intracellular staining with Anti-Mouse/Rat Bcl-2 FITC using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824) and protocol. CD4+ single positive (purple histogram), CD8+ single positive (orange histogram) were analyzed for Bcl-2 expression. Mouse IgG1 K Isotype Control FITC (Product # 11-4714) is shown in the blue histogram.
|Tested species reactivity||Mouse, Rat|
|Host / Isotype||Mouse / IgG1, kappa|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin, 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||5 µL (0.5 µg)/test|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Description: The 10C4 monoclonal antibody reacts with mouse and rat Bcl-2, also known as B cell lymphoma 2. Bcl-2 is the founding member of the Bcl-2 family of evolutionarily-conserved proteins that regulate cell death/survival. Members of the Bcl-2 family include the pro-apoptotic proteins BAD, Bax, and Bim, and the anti-apoptotic proteins Bcl-2, Bcl-xL, and Mcl-1. Bcl-2 inhibits apoptosis by preventing the release of cytochrome c from the mitochondria. Mutation of Bcl-2 or overexpression of Bcl-2 due to chromosomal translocation has been identified as a contributing factor in numerous cancers. In developing thymocytes, Bcl-2 is expressed at high levels in CD4-CD8- double negative and in CD4+ and CD8+ single positive thymocytes; Bcl-2 expression is greatly diminished in CD4+CD8+ double positive thymocytes.
Applications Reported: The 10C4 antibody has been reported for use in intracellular flow cytometric analysis.
Applications Tested: This 10C4 antibody has been pre-titrated and tested by intracellular staining and flow cytometric analysis of mouse splenocytes and thymocytes. This can be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.
Excitation: 488 nm; Emission: 520 nm; Laser: Blue Laser
Members of the Bcl-2 Family (including Bcl-2, Bcl-XLand Bax) play an important role in regulation of apoptosis. Whereas Bcl-2 is commonly regarded as an anti-apoptotic protein, Bax is considered to have a pro-apoptotic function. Regulation of apoptosis is supposed to involve both homo- and heterodimerization of different isoforms of Bax and Bcl-2. The Bax gene encodes different isoforms including Bax alpha (21 kDa) and Bax beta (24 kDa), whereas both isoforms contain the BH1, BH2 and BH3 domains, Bax beta has a unique carboxyl terminus and does not contain a hydrophobic transmembrane domain. Bcl-2 is also expressed in different Isoforms. Bcl-2 beta differs in the 3' UTR and coding region compared to variant alpha. Bcl-2 beta is shorter (22 kDa) and has a distinct C-terminus compared to Bcl-2 alpha (26 kDa).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.