Recombinant Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
![Western blot - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)](https://www.abcam.com/ps/products/218/ab218123/Images/ab218123-306876-anti-bcl-2-phospho-s70-antibody-epr21162-western-blot.jpg "Western blot - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21162] to Bcl-2 (phospho S70)
- Suitable for: Flow Cyt (Intra), WB, Dot blot, ICC/IF, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Bcl-2 (phospho S70) antibody [EPR21162]
See all Bcl-2 primary antibodies -
Description
Rabbit monoclonal [EPR21162] to Bcl-2 (phospho S70) -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, Dot blot, ICC/IF, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Paclitaxel treated Jurkat whole cell lysate. Dot: Bcl-2 (phospho S70) peptide. ICC/IF: HeLa cells. Flow: HeLa cells. IP: Paclitaxel treated Jurkat whole cell lysate.
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General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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
EPR21162 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Corresponding non-phospho antibody
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab218123 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 |
|---|---|---|
| Flow Cyt (Intra) |
1/500.
|
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| WB |
1/1000. Predicted molecular weight: 26 kDa.
|
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| Dot blot |
1/1000.
|
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| ICC/IF |
1/1000.
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| IP |
1/30.
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| Notes |
|---|
|
Flow Cyt (Intra)
1/500. |
|
WB
1/1000. Predicted molecular weight: 26 kDa. |
|
Dot blot
1/1000. |
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ICC/IF
1/1000. |
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IP
1/30. |
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 (phospho S70) antibody [EPR21162] (ab218123)All lanes : Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123) at 1/5000 dilution
Lane 1 : Untreated Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : Jurkat treated with 1 µM paclitaxel for 24 hours whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 26 kDa
Exposure time: 32 secondsBlocking/Dilution buffer and concentration: 2% BSA/TBST
The expression profile observed is consistent with what has been described in the literature (PMID: 10097113).
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![Immunocytochemistry/ Immunofluorescence - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)](https://www.abcam.com/ps/products/218/ab218123/Images/ab218123-306877-anti-bcl-2-phospho-s70-antibody-epr21162-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Bcl-2 (phospho S70) with ab218123 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/100 dilution (green). Confocal image showing positive staining in HeLa cells in M phase. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab195889) at 1/200 dilution (red). Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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![Flow Cytometry (Intracellular) - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)](https://www.abcam.com/ps/products/218/ab218123/Images/ab218123-308017-anti-bcl-2-phospho-s70-antibody-epr21162-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed,90% methanol permeabilized Jurkat (human T cell leukemia T lymphocyte) treated with 1 µM paclitaxel for 24h (Right) / Untreated control (Left) cell line labeling Bcl-2 (phospho S70)with ab218123 at 1/500 dilution. Goat Anti-Rabbit IgG H&L (Alexa Fluor®488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Square gate shows Bcl-2 (phospho S70) positive signal. The expression profile is consistent with what has been described in the literature (PMID: 10097113)
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![Flow Cytometry (Intracellular) - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)](https://www.abcam.com/ps/products/218/ab218123/Images/ab218123-306873-anti-bcl-2-phospho-s70-antibody-epr21162-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)Intracellular flow cytometric analysis of 80% methanol-fixed, 0.1% Tween-20 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling Bcl-2 (phospho S70)with ab218123 at 1/500 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Cells were pretreated with 20 µg/ml RNase A for 30 minutes to eliminate the non-specific binding between RNA and propidium iodide (PI).
Bcl-2 (phospho S70) is highly expressed in mitotic cells (PMID: 10567572).
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![Immunoprecipitation - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)](https://www.abcam.com/ps/products/218/ab218123/Images/ab218123-306874-anti-bcl-2-phospho-s70-antibody-epr21162-immunoprecipitation.jpg)
Immunoprecipitation - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)Bcl-2 (phospho S70) was immunoprecipitated from 0.35 mg of Jurkat (human T cell leukemia cells from peripheral blood) treated with 1 μM paclitaxel for 24 hours whole cell lysate with ab218123 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab218123 at 0.5 μg/ml. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: Jurkat treated with 1 μM paclitaxel for 24 hours whole cell lysate 10 µg (Input).
Lane 2: ab218123 IP in Jurkat treated with 1 μM paclitaxel for 24 hours whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab218123 in Jurkat treated with 1 μM paclitaxel for 24 hours whole cell lysate.
Blocking/ Dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 15 seconds.
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![Dot Blot - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)](https://www.abcam.com/ps/products/218/ab218123/Images/ab218123-306875-anti-bcl-2-phospho-s70-antibody-epr21162-dot-blot.jpg)
Dot Blot - Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)Dot blot analysis of Bcl-2 (phospho S70) labeled with ab218123 at 1/1000 dilution.
Lane 1: Bcl-2 (phospho S70) peptide.
Lane 2: Bcl-2 non-phospho peptide.Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100,000 dilution was used as secondary antibody.
Blocking and dilution buffer: 2% BSA/TBST.
Exposure time: 58 seconds.
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![Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)](https://www.abcam.com/ps/products/218/ab218123/Images/ab218123-7-benefits-of-recombinant-antibodies.png)
Anti-Bcl-2 (phospho S70) antibody [EPR21162] (ab218123)
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
Certificate of Compliance
References (2)
ab218123 has been referenced in 2 publications.
- Wang J et al. Thioredoxin relieves lipopolysaccharide-induced acute kidney injury in mice by reducing inflammation, oxidative stress and apoptosis. Exp Ther Med 21:629 (2021). PubMed: 33936285
- Chu X et al. GSK-J4 induces cell cycle arrest and apoptosis via ER stress and the synergism between GSK-J4 and decitabine in acute myeloid leukemia KG-1a cells. Cancer Cell Int 20:209 (2020). PubMed: 32514253
