Anti-Human BCL2 (Bcl-2/100) ColorWheel^® Dye-Ready mAb

ColorWheel^® technology is a novel and proprietary method of creating your own antibody and dye combinations for use in flow cytometry. By incubating any ColorWheel^® antibody with any ColorWheel^® dye, researchers can quickly and simply produce primary conjugated antibodies for use in single color or multicolor flow cytometry analysis. Each ColorWheel^® antibody and dye is lyophilized for long-term storage, allowing for simplicity and flexibility without compromise. ColorWheel^® technology requires both ColorWheel^® antibody and ColorWheel^® dye for Flow Cytometry application.

Apoptosis regulator Bcl-2 (UniProt: P10415; also known as Bcl2) is encoded by the BCL2 gene (Gene ID: 596) in human. The members of Bcl-2 family of proteins are known to be involved in either blocking or promoting apoptotic cell death. They act at checkpoints upstream of caspases and share regions of homology termed BH (Bcl Homology) domains. All members of the Bcl-2 family (except Bad and Bid) contain a hydrophobic C-terminus (transmembrane domain), which helps them to attach to membrane. Bcl-2 and other anti-apoptotic members of the Bcl-2 family are located in the outer mitochondrial membrane. Bcl-2 is a single-pass membrane protein that is localized in endoplasmic reticulum, nucleus, and mitochondrial membrane. In mitochondria it is especially enriched at contact sites where the inner and outer membranes come in close proximity. It is reported to counteract the action of pore forming Bax protein. Its transmembrane domain is localized in amino acids 212-233. It contains four BH motifs. BH1 (aa 10-30), BH2 (aa 93-107), and BH4 (aa 136-155) are required for its anti-apoptotic activity. Bcl-2 can be present in cells as homodimer or it can form heterodimers with BAX, BAD, BAK and Bcl-X(L). Heterodimerization with BAX requires intact BH1 and BH2 motifs and is necessary for anti-apoptotic activity. Bcl-2 is shown to suppress apoptosis by controlling the mitochondrial membrane permeability. It functions in a feedback loop system with caspases and is shown to inhibit caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF-1). During apoptosis it is proteolytically cleaved by caspases and the cleaved protein lacking the BH4 motif displays pro-apoptotic activity. Following growth factor stimulation, Bcl-2 can undergo phosphorylation at serine 70 by PKC, which is required for its anti-apoptotic activity. (Ref.: Youle, RJ., and Strasser, A. (2008), Nat. Rev. Mol. Cell Biol. 9(1); 47-59; Ruvolo, PP., et al. (2001). Leukemia. 15(4); 515-522).

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A linear peptide corresponding to 14 amino acids from the N-terminal region of human Bcl-2.

Quality Control Testing

Evaluated by Flow Cytometry in Fixed and permeabilized Jurkat cells.

Flow Cytometry Analysis: Staining of one million fixed and permeabilized Jurkat cells was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1145, Anti-Human BCL2 (Bcl-2/100) ColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of Cat. No. CWA-1013 Mouse IgG1 isotype control and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye.

Tested Applications

Flow Cytometry Analysis: Staining of one million fixed and permeabilized human peripheral blood lymphocytes was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1145, Anti-Human BCL2 (Bcl-2/100) ColorWheel^® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheel^® Antibody-Ready Allophycocyanin (APC) Dyeor a 1:1 mixture of an equivalent amount of Cat. No. CWA-101 Mouse IgG1 isotype control and Cat. No. CWDS-APC ColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye.

Flow Cytometry Analysis: Staining of one million fixed and permeabilized human peripheral blood lymphocytes was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1145, Anti-Human BCL2 (Bcl-2/100) ColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of Cat. No. CWA-1013 Mouse IgG1 isotype control and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye.

Flow Cytometry Analysis: Staining of one million fixed and permeabilized Jurkat cells was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1145, Anti-Human BCL2 (Bcl-2/100) ColorWheel^® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye or a 1:1 mixture of an equivalent amount of Cat. No. CWA-101 Mouse IgG1 isotype control and Cat. No. CWDS-APC ColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Clone Bcl2/100 is a mouse monoclonal antibody that detects BCL2. It targets an epitope within 14 amino acids from the N-terminal region.

Lyophilized from PBS containing D-Mannitol and Sucrose, normal appearance is a dried pellet. Reconstituted antibody solution is stable and functional as assessed by functional testing. Contains no biocide or preservatives, such as azide.

1.0 mg/mL after reconstitution at 25 μL in PBS. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommend storage of lyophilized product at 2-8°C. Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial. Reconstitute each vial by adding 25 μL PBS. Protect from light.

ColorWheel is a registered trademark of Merck KGaA, Darmstadt, Germany

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.