Recombinant Anti-Bcr antibody [EPR22062] (ab222406)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22062] to Bcr
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Bcr antibody [EPR22062]
See all Bcr primary antibodies -
Description
Rabbit monoclonal [EPR22062] to Bcr -
Host species
Rabbit -
Specificity
IHC is not recommended for human. -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: K562, PC-12, NIH/3T3, MCF7 and HepG2 whole cell lysates; Human and rat brain tissue lysates; Human testis tissue lysate. IHC-P: Mouse hippocampus and rat cerebrum tissue. Flow Cyt (intra): K562 cells. IP: HeLa 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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22062 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab222406 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 |
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Flow Cyt (Intra) |
1/500.
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WB |
1/1000. Detects a band of approximately 230, 210, 160, 130 kDa (predicted molecular weight: 143 kDa).
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IHC-P |
1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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IP |
1/30.
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Notes |
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Flow Cyt (Intra)
1/500. |
WB
1/1000. Detects a band of approximately 230, 210, 160, 130 kDa (predicted molecular weight: 143 kDa). |
IHC-P
1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
IP
1/30. |
Target
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Function
GTPase-activating protein for RAC1 and CDC42. Promotes the exchange of RAC or CDC42-bound GDP by GTP, thereby activating them. Displays serine/threonine kinase activity. -
Involvement in disease
Note=A chromosomal aberration involving BCR is a cause of chronic myeloid leukemia. Translocation t(9;22)(q34;q11) with ABL1. The translocation produces a BCR-ABL found also in acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). -
Sequence similarities
Contains 1 C2 domain.
Contains 1 DH (DBL-homology) domain.
Contains 1 PH domain.
Contains 1 Rho-GAP domain. -
Domain
The region involved in binding to ABL1 SH2-domain is rich in serine residues and needs to be Ser/Thr phosphorylated prior to SH2 binding. This region is essential for the activation of the ABL1 tyrosine kinase and transforming potential of the chimeric BCR-ABL oncogene.
The DH domain is involved in interaction with CCPG1. -
Post-translational
modificationsAutophosphorylated. Phosphorylated by FES/FPS on tyrosine residues, leading to down-regulation of the BCR kinase activity. Phosphorylation at Tyr-177 by HCK is important for interaction with GRB2. - Information by UniProt
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Database links
- Entrez Gene: 613 Human
- Entrez Gene: 110279 Mouse
- Entrez Gene: 309696 Rat
- Omim: 151410 Human
- SwissProt: P11274 Human
- SwissProt: Q6PAJ1 Mouse
- Unigene: 517461 Human
- Unigene: 715409 Human
see all -
Alternative names
- ALL antibody
- bcr antibody
- BCR/ABL FUSION GENE, INCLUDED antibody
see all
Images
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All lanes : Anti-Bcr antibody [EPR22062] (ab222406) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : BCR CRISPR/Cas9 edited HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 143 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab222406 observed at 150 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab222406 was shown to react with BCR in wild-type HEK-293T cells in western blot. The bands observed in BCR CRISPR/Cas9 edited cell line ab266583 (BCR CRISPR/Cas9 edited cell lysate ab257858) below 150kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and BCR CRISPR/Cas9 edited HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab222406 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4
176;
® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging. -
Bcr was immunoprecipitated from 0.35 mg HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab222406 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab222406 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection antibody at 1/1000 dilution.
Lane 1: HeLa whole cell lysate 10μg (Input).
Lane 2: ab222406 IP in HeLa whole cell lysate (+)
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab222406 in HeLa whole cell lysate (-).Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.The 160 kDa and 130 kDa bands are isoforms of BCR (PMID:2494632, PMID:3078961).
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized K562 (human chronic myelogenous leukemia cell line from bone marrow) cell line labeling Bcr with ab222406 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
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Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling Bcr with ab222406 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use. Cytoplasmic staining in rat cerebrum (PMID: 25331951). Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use.
Heat-mediated antigen retrieval using ab208572 (Universal HIER antigen retrieval reagent).
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Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling Bcr with ab222406 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use. Cytoplasmic staining in mouse hippocampus (PMID: 25331951). Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use.
Heat-mediated antigen retrieval using ab208572 (Universal HIER antigen retrieval reagent).
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All lanes : Anti-Bcr antibody [EPR22062] (ab222406) at 1/1000 dilution
Lane 1 : MCF7 (human breast adenocarcinoma cell line) whole lysate
Lane 2 : Human brain lysate
Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) whole lysate
Lane 4 : Rat brain lysate
Lane 5 : Human testis 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: 143 kDa
Observed band size: 130,160 kDa why is the actual band size different from the predicted?Exposure time : Lanes 1 and 2: 3 minutes; Lanes 3 and 4: 59 seconds; Lane 3: 81 seconds.
Blocking/Dilution buffer: 5% NFDM/TBST.
The 160 kDa and 130 kDa bands are isoforms of BCR (PMID: 2494632, PMID: 3078961).
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All lanes : Anti-Bcr antibody [EPR22062] (ab222406) at 1/1000 dilution
Lane 1 : K562 (human chronic myelogenous leukemia cell line from bone marrow) whole lysate
Lane 2 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole lysate
Lane 3 : NIH/3T3 (mouse embyro fibroblast cell line) whole 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: 143 kDa
Observed band size: 130,160,210,230 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
The 230/210 kDa band is the BCR-ABL fusion protein in K562 cells (PMID:17684099, PMID:16585609, PMID:12476301). The 160 kDa and 130 kDa bands are isoforms of BCR (PMID:2494632, PMID:3078961).
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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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab222406 has not yet been referenced specifically in any publications.