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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, Ca, Fe | FC, IH(P), WB | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 05-729 |
| Replaces | 04-436 |
| Brand Family | Upstate |
| Trade Name |
|
| Description | Anti-Bcl2 Antibody, clone 100 |
| Alternate Names |
|
| Background Information | 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. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
|
| Presentation | Purified mouse monoclonal IgG1 in buffer containing 70% storage buffer PBS with 0.1% sodium azide and 30% glycerol. Store at -20ºC. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Detect Bcl2 using this Anti-Bcl2 Antibody, clone 100 validated for use in FC, IH & WB. |
| Key Applications |
|
| Application Notes | Flow Cytometry analysis: A representative lot of this antibody detected Bcl2 in Jurkat cells. Western Blotting Analysis: 1 µg/mL of this antibody detected Bcl2 in Raji cell lysate. Immunohistochemistry(Paraffin) Analysis: A 1:1,000 dilution of this antibody detected Bcl2 in Human tonsil tissue sections. |
| Biological Information | |
|---|---|
| Immunogen | Synthetic peptide corresponding to amino acids 41-54 of human Bcl2 protein. The immunizing sequence is identical in canine and feline. |
| Epitope | a.a. 41-54 |
| Clone | 100 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | This antibody recognizes Bcl2, Mr 26 kDa. |
| Isotype | IgG1 |
| Species Reactivity |
|
| Species Reactivity Note | Human. Predicted to cross-react with feline and canine, based on sequence homology. |
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | This gene encodes an integral outer mitochondrial membrane protein that blocks the apoptotic death of some cells such as lymphocytes. 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. Two transcript variants, produced by alternate splicing, differ in their C-terminal ends. |
| Gene Symbol |
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| Purification Method | Protein G Purified |
| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: P10415 # Suppresses apoptosis in a variety of cell systems including factor-dependent lymphohematopoietic and neural cells. Regulates cell death by controlling the mitochondrial membrane permeability. Appears to function in a feedback loop system with caspases. Inhibits caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF-1). SIZE: 239 amino acids; 26266 Da SUBUNIT: Forms homodimers, and 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 (By similarity). Also interacts with APAF1, RAF-1, TP53BP2, BBC3, BCL2L1, MRPL41 and BNIPL. Binding to FKBP8 seems to target BCL2 to the mitochondria and probably interferes with the binding of BCL2 to its targets. SUBCELLULAR LOCATION: Mitochondrion outer membrane; Single-pass membrane protein. Nucleus membrane; Single-pass membrane protein. Endoplasmic reticulum membrane; Single-pass membrane protein. TISSUE SPECIFICITY: Expressed in a variety of tissues. DOMAIN: SwissProt: P10415 The BH4 motif is required for anti-apoptotic activity and for interaction with RAF-1. PTM: Phosphorylation/dephosphorylation on Ser-70 regulates anti- apoptotic activity. Growth factor-stimulated phosphorylation on Ser-70 by PKC is required for the anti-apoptosis activity and occurs during the G2/M phase of the cell cycle. In the absence of growth factors, BCL2 appears to be phosphorylated by other protein kinases such as ERKs and stress-activated kinases. Dephosphorylated by protein phosphatase 2A (PP2A) (By similarity). & Proteolytically cleaved by caspases during apoptosis. The cleaved protein, lacking the BH4 motif, has pro-apoptotic activity, causes the release of cytochrome c into the cytosol promoting further caspase activity. DISEASE: SwissProt: P10415 # A chromosomal aberration involving BCL2 may be a cause of follicular lymphoma (FL) [MIM:151430]; also known as type II chronic lymphatic leukemia. Translocation t(14;18)(q32;q21) with immunoglobulin gene regions. BCL2 mutations found in non-Hodgkin lymphomas carrying the chromosomal translocation could be attributed to the Ig somatic hypermutation mechanism resulting in nucleotide transitions. SIMILARITY: Belongs to the Bcl-2 family. |
| Molecular Weight | 26 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Routinely evaluated by western blot on Raji RIPA cell lysate. Western Blot Analysis: 0.5-2 µg/mL of this lot detected Bcl2 in a Raji RIPA cell lysate. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20ºC. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20ºC may cause glycerol-containing solutions to become frozen during storage. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |