Rabbit Bcl-2 Polyclonal Antibody | anti-Bcl-2 antibody

Phospho-Bcl-2 (Thr74) Antibody

Cat. Num. AAA9614458

• Gene Names: BCL2; Bcl-2; PPP1R50
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Horse (100%), Dog (86%)
• Applications: Western Blot, Immunohistochemistry, Immunofluorescence, Immunocytochemistry, ELISA
• Purity: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Synonyms: Bcl-2; Polyclonal Antibody; Phospho-Bcl-2 (Thr74) Antibody; Apoptosis regulator Bcl 2; Apoptosis regulator Bcl-2; Apoptosis regulator Bcl2; AW986256; B cell CLL/lymphoma 2; B cell leukemia/lymphoma 2; Bcl2; BCL2_HUMAN; C430015F12Rik; D630044D05Rik; D830018M01Rik; Leukemia/lymphoma; B-cell; 2; Oncogene B-cell leukemia 2; PPP1R50; Protein phosphatase 1; regulatory subunit 50; anti-Bcl-2 antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Horse (100%), Dog (86%)
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Phospho-Bcl-2 (Thr74) Antibody detects endogenous levels of Bcl-2 only when phosphorylated at Thr74.; Tissue Specificity: Expressed in a variety of tissues.
• Purity/Purification: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Form/Format: Liquid. Rabbit IgG in phosphate buffered saline, pH7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
• Concentration: 1mg/ml (varies by lot)

• Applicable Applications for anti-Bcl-2 antibody: Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
• Application Notes: WB: 1:1000-3000; IF/ICC: 1:100-1:500; IHC: 1:50-1:200; Peptide ELISA: 1:20,000-1:40,000
• Immunogen: A synthesized peptide derived from human Bcl-2 around the phosphorylation site of Thr74.
• Conjugation: Unconjugated
• Fragment: Fab fragment
• Post Translational Modifications: 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. Phosphorylated by MAPK8/JNK1 at Thr-69, Ser-70 and Ser-87, wich stimulates starvation-induced autophagy. 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.Monoubiquitinated by PRKN, leading to increase its stability. Ubiquitinated by SCF (FBXO10), leading to its degradation by the proteasome.
• Subunit Structure: 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. Interacts with EI24 (By similarity). Also interacts with APAF1, BBC3, BCL2L1, BNIPL, MRPL41 and TP53BP2. Binding to FKBP8 seems to target BCL2 to the mitochondria and probably interferes with the binding of BCL2 to its targets. Interacts with BAG1 in an ATP-dependent manner. Interacts with RAF1 (the 'Ser-338' and 'Ser-339' phosphorylated form). Interacts (via the BH4 domain) with EGLN3; the interaction prevents the formation of the BAX-BCL2 complex and inhibits the anti-apoptotic activity of BCL2. Interacts with G0S2; this interaction also prevents the formation of the anti-apoptotic BAX-BCL2 complex. Interacts with RTL10/BOP. Interacts with the SCF (FBXO10) complex. Interacts (via the loop between motifs BH4 and BH3) with NLRP1 (via LRR repeats), but not with NLRP2, NLRP3, NLRP4, PYCARD, nor MEFV. Interacts with GIMAP3/IAN4, GIMAP4/IAN1 and GIMAP5/IAN5 (By similarity).
• Similarity: BH1 and BH2 domains are required for the interaction with BAX and for anti-apoptotic activity.The BH4 motif is required for anti-apoptotic activity and for interaction with RAF1 and EGLN3.The loop between motifs BH4 and BH3 is required for the interaction with NLRP1.Belongs to the Bcl-2 family.
• Subcellular Location: Mitochondrion outer membrane>Single-pass membrane protein. Nucleus membrane>Single-pass membrane protein. Endoplasmic reticulum membrane>Single-pass membrane protein.
• Preparation and Storage: Store at -20 degree C. Stable for 12 months from date of receipt.

Western Blot (WB)

(Western blot analysis of BCL-2 (Phospho-Thr74) using EGF treated HepG2 whole cell lysates.-/+ means absence or presence of N peptide（non-phospho peptide) and P peptide(phospho peptide).)

Immunohistochemistry (IHC)

(Staining Rat brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Mouse testis tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Mouse brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Testing Data

(At 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)

Related Product Information for anti-Bcl-2 antibody

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). May attenuate inflammation by impairing NLRP1-inflammasome activation, hence CASP1 activation and IL1B release.

NCBI and Uniprot Product Information

• NCBI GI #: 72198189
• NCBI GeneID: 596
• NCBI Accession #: NP_000624.2
• NCBI GenBank Nucleotide #: NM_000633.2
• UniProt Accession #: P10415; P10416; Q13842; Q16197
• Molecular Weight: 26,266 Da
• NCBI Official Full Name: apoptosis regulator Bcl-2 alpha isoform
• NCBI Official Synonym Full Names: B-cell CLL/lymphoma 2
• NCBI Official Symbol: BCL2
• NCBI Official Synonym Symbols: Bcl-2; PPP1R50
• NCBI Protein Information: apoptosis regulator Bcl-2; protein phosphatase 1, regulatory subunit 50
• UniProt Protein Name: Apoptosis regulator Bcl-2
• Protein Family: Bcl-2-like protein
• UniProt Gene Name: BCL2
• UniProt Entry Name: BCL2_HUMAN

NCBI Description

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. [provided by RefSeq, Jul 2008]

Uniprot Description

Bcl-2: a antiapoptotic member of the Bcl-2 family. Regulates cell death by controlling the mitochondrial membrane permeability. 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). Phosphorylation by JNKs may increase its antiapoptotic functions.

Protein type: Membrane protein, integral; Oncoprotein; Autophagy; Apoptosis

Chromosomal Location of Human Ortholog: 18q21.3

Cellular Component: pore complex; endoplasmic reticulum membrane; mitochondrial outer membrane; nuclear membrane; membrane; mitochondrion; endoplasmic reticulum; cytoplasm; nucleus; cytosol; myelin sheath

Molecular Function: identical protein binding; protein binding; protein homodimerization activity; protease binding; protein phosphatase 2A binding; sequence-specific DNA binding; channel activity; protein heterodimerization activity; ubiquitin protein ligase binding; BH3 domain binding; channel inhibitor activity; transcription factor binding

Biological Process: focal adhesion formation; response to nicotine; positive regulation of catalytic activity; developmental growth; renal system process; pigment granule organization and biogenesis; protein polyubiquitination; response to toxin; response to glucocorticoid stimulus; T cell differentiation in the thymus; ear development; lymphoid progenitor cell differentiation; female pregnancy; positive regulation of multicellular organism growth; glomerulus development; negative regulation of mitochondrial depolarization; post-embryonic development; cochlear nucleus development; cellular response to glucose starvation; B cell receptor signaling pathway; negative regulation of myeloid cell apoptosis; regulation of mitochondrial membrane potential; negative regulation of ossification; positive regulation of B cell proliferation; regulation of transmembrane transporter activity; T cell homeostasis; negative regulation of neuron apoptosis; cell growth; defense response to virus; spleen development; response to drug; positive regulation of neuron maturation; release of cytochrome c from mitochondria; axon regeneration; regulation of protein homodimerization activity; actin filament organization; cell aging; digestive tract morphogenesis; regulation of calcium ion transport; positive regulation of cell growth; organ growth; induction of apoptosis via death domain receptors; DNA damage response, signal transduction resulting in induction of apoptosis; negative regulation of osteoblast proliferation; gland morphogenesis; regulation of mitochondrial membrane permeability; regulation of nitrogen utilization; metanephros development; oocyte development; negative regulation of apoptosis; B cell proliferation; negative regulation of autophagy; regulation of protein heterodimerization activity; behavioral fear response; melanin metabolic process; apoptosis; regulation of cell-matrix adhesion; negative regulation of retinal cell programmed cell death; regulation of protein stability; protein amino acid dephosphorylation; positive regulation of smooth muscle cell migration; response to radiation; B cell homeostasis; ovarian follicle development; positive regulation of skeletal muscle fiber development; positive regulation of melanocyte differentiation; melanocyte differentiation; response to gamma radiation; negative regulation of cell migration; regulation of viral genome replication; response to iron ion; transmembrane transport; negative regulation of cellular pH reduction; mesenchymal cell development; ossification; hair follicle morphogenesis; CD8-positive, alpha-beta T cell lineage commitment; thymus development; B cell lineage commitment; male gonad development; peptidyl-threonine phosphorylation; positive regulation of peptidyl-serine phosphorylation; humoral immune response; response to UV-B; endoplasmic reticulum calcium ion homeostasis; neuron apoptosis; peptidyl-serine phosphorylation; response to hydrogen peroxide; axonogenesis; ureteric bud branching; homeostasis of number of cells within a tissue; response to cytokine stimulus; innate immune response; negative regulation of cell growth; response to DNA damage stimulus; induction of apoptosis by oxidative stress