Anti-Bcl-2 antibody [EPR17509] (ab182858)
- Datasheet
- References (24)
- Protocols
Overview
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Product nameAnti-Bcl-2 antibody [EPR17509]
See all Bcl-2 primary antibodies -
DescriptionRabbit monoclonal [EPR17509] to Bcl-2
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Host speciesRabbit
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Tested applicationsSuitable for: WB, ICC/IF, Flow Cyt, IHC-Pmore details
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Species reactivityReacts with: Mouse, Human
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Immunogen
Recombinant fragment within Human Bcl-2 aa 1 to the C-terminus. The exact sequence is proprietary.
Database link: P10415 -
Positive control
- WB: Human tonsil lysate, Human thymus lysate; Jurkat, U-937, THP-1, HeLa, C2C12, WEHI -3 and NIH/3T3 whole cell lysates; Mouse brain lysate, Mouse heart lysate, Mouse kidney lysate, Mouse spleen lysate, Human fetal kidney lysate, Human fetal spleen lysate. IHC-P: Human tonsil tissue, Human endometrial cancer tissue, Mouse spleen tissue. ICC/IF: Jurkat cells. Flow Cyt: Jurkat cells.
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General notes
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
This product is a recombinant rabbit monoclonal antibody.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
PurityProtein A purified
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ClonalityMonoclonal
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Clone numberEPR17509
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IsotypeIgG
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Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
Applications
Our Abpromise guarantee covers the use of ab182858 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | 1/2000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa). | |
| ICC/IF | 1/150. | |
| Flow Cyt | 1/250. | |
| IHC-P | 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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FunctionSuppresses 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).
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Tissue specificityExpressed in a variety of tissues.
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Involvement in diseaseA chromosomal aberration involving BCL2 has been found in 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.
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Sequence similaritiesBelongs to the Bcl-2 family.
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DomainBH1 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. -
Post-translational
modificationsPhosphorylation/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).
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 PARK2, leading to increase its stability. Ubiquitinated by SCF(FBXO10), leading to its degradation by the proteasome. -
Cellular localizationMitochondrion outer membrane. Nucleus membrane. Endoplasmic reticulum membrane.
- Information by UniProt
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Database links
- Entrez Gene: 596 Human
- Entrez Gene: 12043 Mouse
- Omim: 151430 Human
- SwissProt: P10415 Human
- SwissProt: P10417 Mouse
- Unigene: 150749 Human
- Unigene: 257460 Mouse
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Alternative names
- Apoptosis regulator Bcl 2 antibody
- Apoptosis regulator Bcl-2 antibody
- Apoptosis regulator Bcl2 antibody
see all
Images
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![Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)](https://www.abcam.com/ps/products/182/ab182858/Images/ab182858-275407-182858KOWB.jpg)
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: BCL2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: THP-1 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab182858 observed at 26 kDa. Red - loading control, ab8245, observed at 37 kDa.ab182858 was shown to specifically react with BCL2 when BCL2 knockout samples were used. Wild-type and BCL2 knockout samples were subjected to SDS-PAGE. Ab182858 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)](https://www.abcam.com/ps/products/182/ab182858/Images/ab182858-245900-Bcl2IHC1.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Bcl-2 with ab182858 at 1/1000 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.
Cytoplasm, nuclear membrane and nucleus staining on lymphocytes of Human tonsil tissue is observed.
Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody followed by ab97051 at 1/500.
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![Flow Cytometry - Anti-Bcl-2 antibody [EPR17509] (ab182858)](https://www.abcam.com/ps/products/182/ab182858/Images/ab182858-245904-Bcl2FC.jpg)
Flow Cytometry - Anti-Bcl-2 antibody [EPR17509] (ab182858)Flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Bcl-2 with ab182858 at 1/250 (red) compared with a rabbit monoclonal IgG isotype control (ab172730) (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody (blue)). Goat anti rabbit IgG (FITC) at 1/500 was used as the secondary antibody.
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![Immunocytochemistry/ Immunofluorescence - Anti-Bcl-2 antibody [EPR17509] (ab182858)](https://www.abcam.com/ps/products/182/ab182858/Images/ab182858-245903-BCl2IF1.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Bcl-2 antibody [EPR17509] (ab182858)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Bcl-2 with ab182858 at 1/150, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 (green).
Confocal image showing cytoplasmic and weak nuclear staining on Jurkat cells line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 (red).
The negative controls are as follows:-
-ve control 1 - ab182858 at 1/150 followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000. -
![Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)](https://www.abcam.com/ps/products/182/ab182858/Images/ab182858-262061-182858WB1.jpg)
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/10000 dilution
Lane 1 : NIH/3T3 (Mouse fibroblast) whole cell lysate at 20 µg
Lane 2 : WEHI-3 (Mouse myelomonocyte from leukemia) whole cell lysate at 20 µg
Lane 3 : Mouse hippocampus at 10 µg
Lane 4 : Mouse heart at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) at 1/2000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDa
Exposure time: 8 secondsBlocking/Diluting buffer 5% NFDM/TBST
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)](https://www.abcam.com/ps/products/182/ab182858/Images/ab182858-245901-Bcl2IHC2.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labeling Bcl-2 with ab182858 at 1/1000 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.
Cytoplasm, nuclear membrane and nucleus staining on lymphocytes and cancer cells of Human endometrial cancer tissue is observed.
Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody followed by ab97051 at 1/500. -
![Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)](https://www.abcam.com/ps/products/182/ab182858/Images/ab182858-245897-Bcl2WB1.jpg)
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/20000 dilution
Lane 1 : Human tonsil lysate
Lane 2 : Human thymus lysate
Lane 3 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lane 4 : U-937 (Human histiocytic lymphoma cells) whole cell lysate
Lane 5 : THP-1 (Human monocytic leukemia cells) whole cell lysate
Lane 6 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 7 : C2C12 (Mouse myoblast cell line) whole cell lysate
Lane 8 : WEHI-3 (Mouse leukemia cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 26 kDa
Observed band size: 26 kDa
Exposure time: 1 minuteBlocking and diluting buffer was 5% NFDM /TBST.
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![Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)](https://www.abcam.com/ps/products/182/ab182858/Images/ab182858-245898-Bcl2WB2.jpg)
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/2000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Mouse spleen lysate
Lane 5 : NIH/3T3 (Mouse embryo fibroblast cells) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 26 kDa
Observed band size: 26 kDa
Exposure time: 3 minutesBlocking and diluting buffer was 5% NFDM /TBST.
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![Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)](https://www.abcam.com/ps/products/182/ab182858/Images/ab182858-245899-Bcl2WB3.jpg)
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/2000 dilution
Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal spleen lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 26 kDa
Observed band size: 26 kDa
Exposure time: 3 minutesBlocking and diluting buffer was 5% NFDM /TBST.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)](https://www.abcam.com/ps/products/182/ab182858/Images/ab182858-245902-Bcl2IHC3.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Bcl-2 with ab182858 at 1/1000 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.
Cytoplasm, nuclear membrane and nucleus staining on lymphocytes of Mouse spleen tissue is observed.
Counter stained with Hematoxylin.
Negative control: Used PBS instead of primary antibody followed by ab97051 at 1/500.
Protocols
Datasheets and documents
References
This product has been referenced in:
- Zhao W et al. Long noncoding RNA NSCLCAT1 increases non-small cell lung cancer cell invasion and migration through the Hippo signaling pathway by interacting with CDH1. FASEB J 33:1151-1166 (2019). Read more (PubMed: 30148675) »
- Hao L et al. Upregulated circular RNA circ_0007534 indicates an unfavorable prognosis in pancreatic ductal adenocarcinoma and regulates cell proliferation, apoptosis, and invasion by sponging miR-625 and miR-892b. J Cell Biochem 120:3780-3789 (2019). Read more (PubMed: 30382592) »
