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Immunofluorescence (IF)
(MBS9600911 staining Jurkat cells by ICC/IF. Cells were fixed with PFA and permeabilized in 0.1% saponin prior to blocking in 10% serum for 45 minutes at 37 degree C. The primary antibody was diluted 1/400 and incubated with the sample for 1 hour at 37 degree C. A Alexa Fluor 594 conjugated goat polyclonal to rabbit IgG (H+L), diluted 1/600 was used as secondary antibody.)
Rabbit BCL-2 Polyclonal Antibody | anti-BCL-2 antibody
Phospho-BCL-2 (Thr69) Antibody
Gene Names
BCL2; Bcl-2; PPP1R50
Reactivity
Human, Mouse, Rat
Applications
Western Blot, Immunohistochemistry, Immunofluorescence, Immunocytochemistry, ELISA
Purity
From purified rabbit serum by affinity purification via sequential chromatography on phospho-and non-phospho-peptide affinity columns.
Synonyms
BCL-2; Polyclonal Antibody; Phospho-BCL-2 (Thr69) Antibody; Apoptosis regulator Bcl 2; Apoptosis regulator Bcl-2; Apoptosis regulator Bcl2; AW986256; B cell CLL/lymphoma 2; B cell leukemia/lymphoma 2; Bcl-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
Clonality
Polyclonal
Isotype
IgG
Specificity
Phospho-BCL-2 (Thr69) antibody detects endogenous levels of BCL-2 only when phosphorylated at Threonine 69
Purity/Purification
From purified rabbit serum by affinity purification via sequential chromatography on phospho-and non-phospho-peptide affinity columns.
Form/Format
Liquid
Phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Concentration
1mg/ml (varies by lot)
Sequence Length
239
Applicable Applications for anti-BCL-2 antibody
Western Blot (WB), Immunohistochemisty (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC), ELISA (EIA)
Application Notes
WB: 1:500
IHC: 1:50-1:200
IF/ICC: 1:100-1:500
IHC: 1:50-1:200
IF/ICC: 1:100-1:500
Immunogen
A synthesized peptide derived from human BCL-2 around the phosphorylation site of Threonine 69
Subcellular Location
Mitochondrion Outer Membrane. Nucleus Membrane. Endoplasmic Reticulum Membrane.
Tissue Specificity
Expressed in a variety of tissues.
Predicted Cross Reactivity
Horse, Dog
Similarity
Horse (100%), Dog (86%)
Conjugation
Unconjugated
Epitope
Phospho Thr69
Preparation and Storage
Store at -20 degree C. Stable for 12 months from date of receipt.
Immunofluorescence (IF)
(MBS9600911 staining Jurkat cells by ICC/IF. Cells were fixed with PFA and permeabilized in 0.1% saponin prior to blocking in 10% serum for 45 minutes at 37 degree C. The primary antibody was diluted 1/400 and incubated with the sample for 1 hour at 37 degree C. A Alexa Fluor 594 conjugated goat polyclonal to rabbit IgG (H+L), diluted 1/600 was used as secondary antibody.)
Immunofluorescence (IF)
(MBS9600911 staining Jurkat cells by ICC/IF. Cells were fixed with PFA and permeabilized in 0.1% saponin prior to blocking in 10% serum for 45 minutes at 37 degree C. The primary antibody was diluted 1/400 and incubated with the sample for 1 hour at 37 degree C. A Alexa Fluor 594 conjugated goat polyclonal to rabbit IgG (H+L), diluted 1/600 was used as secondary antibody.)
Immunofluorescene (IF)
(MBS9600911 staining MCF7 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes 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.)
Immunofluorescene (IF)
(MBS9600911 staining MCF7 by IF/ICC. The sample were fixed with PFA and permeabilized in 0.1% Triton X-100, then blocked in 10% serum for 45 minutes 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.)
Immunohistochemistry (IHC)
(MBS9600911 at 1/200 staining human kidney tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
Immunohistochemistry (IHC)
(MBS9600911 at 1/200 staining human kidney tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.)
Western Blot (WB)
(Western blot analysis of BCL-2 phosphorylation expression in mouse muscle whole tissue lysates, The lane on the right is treated with the antigen-specific peptide.)
Western Blot (WB)
(Western blot analysis of BCL-2 phosphorylation expression in mouse muscle whole tissue lysates, The lane on the right is treated with the antigen-specific peptide.)
Related Product Information for anti-BCL-2 antibody
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.
Function: 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 (PubMed:17418785).
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 (PubMed:8183370). 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 (PubMed:17418785).
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.
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.
Function: 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 (PubMed:17418785).
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 (PubMed:8183370). 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 (PubMed:17418785).
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.
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.
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
Molecular Weight
Observed: 28 kDa
Predicted: 27 kDa
Predicted: 27 kDa
NCBI Official Full Name
apoptosis regulator Bcl-2 isoform alpha
NCBI Official Synonym Full Names
BCL2, apoptosis regulator
NCBI Official Symbol
BCL2
NCBI Official Synonym Symbols
Bcl-2; PPP1R50
NCBI Protein Information
apoptosis regulator Bcl-2
UniProt Protein Name
Apoptosis regulator Bcl-2
Protein Family
UniProt Gene Name
BCL2
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. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Feb 2016]
Uniprot Description
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 (PubMed:17418785).
Research Articles on BCL-2
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Product Notes
The BCL-2 bcl2 (Catalog #AAA9600911) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Phospho-BCL-2 (Thr69) Antibody reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's BCL-2 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemisty (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC), ELISA (EIA). WB: 1:500 IHC: 1:50-1:200 IF/ICC: 1:100-1:500. Researchers should empirically determine the suitability of the BCL-2 bcl2 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "BCL-2, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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