Recombinant Anti-Bcl-2 antibody [NOR 235J] (ab241548)
![Western blot - Anti-Bcl-2 antibody [NOR 235J] (ab241548)](https://www.abcam.com/ps/products/241/ab241548/Images/ab241548-483982-AntiBcl2-antibody-NOR-235J-WB-human.jpg "Western blot - Anti-Bcl-2 antibody [NOR 235J] (ab241548)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rat monoclonal [NOR 235J] to Bcl-2
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Bcl-2 antibody [NOR 235J]
See all Bcl-2 primary antibodies -
Description
Rat monoclonal [NOR 235J] to Bcl-2 -
Host species
Rat -
Tested applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant full length protein. This information is considered to be commercially sensitive.
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Positive control
- IHC-P: Human tonsil tissue. WB: Human spleen tissue, THP-1 cell line.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Properties
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Form
Liquid -
Storage instructions
Shipped 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. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
NOR 235J -
Isotype
IgG2b -
Research areas
Associated products
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Alternative Versions
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab241548 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 |
|---|---|---|
| IHC-P |
Use a concentration of 0.98 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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| WB |
1/1000. Predicted molecular weight: 26 kDa.
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| Notes |
|---|
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IHC-P
Use a concentration of 0.98 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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WB
1/1000. Predicted molecular weight: 26 kDa. |
Target
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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). -
Tissue specificity
Expressed in a variety of tissues. -
Involvement in disease
A 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. -
Sequence similarities
Belongs to the Bcl-2 family. -
Domain
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. -
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 localization
Mitochondrion outer membrane. Nucleus membrane. Endoplasmic reticulum membrane. - Information by UniProt
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Database links
- Entrez Gene: 596 Human
- Omim: 151430 Human
- SwissProt: P10415 Human
- Unigene: 150749 Human
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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 [NOR 235J] (ab241548)All lanes : Anti-Bcl-2 antibody [NOR 235J] (ab241548) at 1/1000 dilution
Lane 1 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lane 2 : U937 (human histiocytic lymphoma monocyte), Whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 4 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate
Lane 5 : Human tonsil tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/10000 dilution (Goat Anti-Rat IgG (H+L), HRP))
Predicted band size: 26 kDa
Observed band size: 26 kDaExposure time:
Lane 1,2,5:3 seconds
Lane 3,4:10 seconds
Blocking buffer / Diluting buffer and concentration:
5% NFDM/TBST
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![Western blot - Anti-Bcl-2 antibody [NOR 235J] (ab241548)](https://www.abcam.com/ps/products/241/ab241548/Images/ab241548-5-ab241548R16annotated.jpg)
Western blot - Anti-Bcl-2 antibody [NOR 235J] (ab241548)All lanes : Anti-Bcl-2 antibody [NOR 235J] (ab241548) at 0.5 µg/ml
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : BCL2 knockout HAP1 whole cell lysate
Lane 3 : NIH/3T3 whole cell lysate
Lane 4 : THP-1 whole cell lysate
Lane 5 : Human Spleen tissue lysate
Lane 6 : Mouse Spleen tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rat IgG H&L (IRDye® 800CW ) (ab253031) at 1/20000 dilution
Predicted band size: 26 kDaLanes 1 - 6: Merged signal (red and green). Green - ab241548 observed at 26 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.
ab241548 was shown to react with Bcl-2 in Western blot. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween) before incubation with ab241548 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at 0.5 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat Anti-Rat IgG H&L (IRDye 800CW) (ab253031) and Goat anti-Rabbit IgG H&L (IRDye 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [NOR 235J] (ab241548)](https://www.abcam.com/ps/products/241/ab241548/Images/ab241548-357746-antibcl2nor235jimmunohistochemistrytonsilhuman.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [NOR 235J] (ab241548)IHC image of Bcl-2 staining in a section of formalin-fixed paraffin-embedded normal human tonsil* performed on a Leica BOND™ system using the standard Protocol F.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab241548, 0.98 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX.
The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.
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![Anti-Bcl-2 antibody [NOR 235J] (ab241548)](https://www.abcam.com/ps/products/241/ab241548/Images/ab241548-3-benefits-of-recombinant-antibodies.png)
Anti-Bcl-2 antibody [NOR 235J] (ab241548)
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (0)
ab241548 has not yet been referenced specifically in any publications.
