Invitrogen™ Bcl-2 Monoclonal Antibody (10C4), eFluor™ 660, eBioscience™

Catalog No.	Quantity
50-699-242	100 Tests

Catalog No. 50-699-242 Supplier Invitrogen™ Supplier No. 50699242

Description

Mouse Monoclonal Antibody

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: This 10C4 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This 10C4 antibody has been pre-titrated and tested by intracellular flow cytometric analysis of mouse splenocytes and thymocytes using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Please refer to BestProtocols®: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This can be used at 5 μL (0.25 μg) per test. A test is defined as the amount (μg) of antibody that will stain a cell sample in a final...

BCL-2 is a key regulator of apoptosis that functions to either inhibit or promote cell death. The BCL-2 family members are also characterized by dimerizing to further modulate apoptosis. Bag1, for example, has been found to form a heterodimer with BCL-2 resulting in the enhancement of the anti-apoptotic effect of BCL-2. Bax and Bak have been shown to play a critical role in cytochrome c release from mitochondria and thus initiate apoptosis. Bax exerts a pro-apoptotic rather than an anti-apoptotic effect on cells. Constitutive expression of BCL2, such as in the case of translocation of BCL2 to Ig heavy chain locus, is thought to be the cause of follicular lymphoma. In most follicular lymphomas, neoplastic germinal centers express high levels of BCL-2 alpha protein, whereas the normal or hyperplastic germinal centers are negative. Two transcript variants of BCL-2, produced by alternate splicing, differ in their C-terminal ends. The overexpression of BCL-2 has been linked to human cancers such as B-cell lymphoma and prostate cancer.

Specifications

• Antigen: Bcl-2
• Applications: Flow Cytometry
• Classification: Monoclonal
• Clone: 10C4
• Concentration: 5 μL/Test
• Conjugate: eFluor 660
• Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
• Gene: Bcl2
• Gene Accession No.: P10417, P49950
• Gene Alias: apoptosis regulator Bcl-2; AW986256; B cell leukemia/lymphoma 2; B cell lymphoma 2 associated oncogene; B-cell CLL/lymphoma 2; B-cell leukemia/lymphoma 2; Bcl 2; BCL2; bcl-2; BCL2 apoptosis regulator; Bcl-2 protein; BCL2, apoptosis regulator; Bcl2-like protein; C430015F12Rik; D630044D05Rik; D830018M01Rik; OTTHUMP00000163680; PPP1R50; protein phosphatase 1, regulatory subunit 50
• Gene Symbols: Bcl2
• Host Species: Mouse
• Purification Method: Affinity chromatography
• Quantity: 100 Tests
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 12043, 24224
• Target Species: Mouse, Rat
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG1 κ

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

Is the eFluor 660 fluorochrome compatible with Anti-Cy5/Alexa Fluor 647 beads?

Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

Are the eFluor Organic Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.