RecombinantRabMAb

Recombinant Anti-Bcr antibody [EPR22062] (ab222406)

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Western blot - Anti-Bcr antibody [EPR22062] (ab222406)
  • Immunoprecipitation - Anti-Bcr antibody [EPR22062] (ab222406)
  • Flow Cytometry (Intracellular) - Anti-Bcr antibody [EPR22062] (ab222406)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcr antibody [EPR22062] (ab222406)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcr antibody [EPR22062] (ab222406)
  • Western blot - Anti-Bcr antibody [EPR22062] (ab222406)
  • Western blot - Anti-Bcr antibody [EPR22062] (ab222406)
  • 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

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • 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.

  • 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.

  • 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
    For more information see here.

    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

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
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.
Notes
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

  • 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
    modifications

    Autophosphorylated. 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

  • Database links

    see all

  • Alternative names

    • ALL antibody
    • bcr antibody
    • BCR/ABL FUSION GENE, INCLUDED antibody
    • BCR/FGFR1 chimera protein antibody
    • BCR/FGFR1 FUSION GENE, INCLUDED antibody
    • BCR/PDGFRA FUSION GENE, INCLUDED antibody
    • BCR_HUMAN antibody
    • BCR1 antibody
    • Breakpoint cluster region antibody
    • Breakpoint cluster region protein antibody
    • CML antibody
    • D22S11 antibody
    • D22S662 antibody
    • FGFR1/BCR chimera protein antibody
    • PHL antibody
    • Renal carcinoma antigen NY-REN-26 antibody
    see all

Images

  • Western blot - Anti-Bcr antibody [EPR22062] (ab222406)
    Western blot - Anti-Bcr antibody [EPR22062] (ab222406)
    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.

  • Immunoprecipitation - Anti-Bcr antibody [EPR22062] (ab222406)
    Immunoprecipitation - Anti-Bcr antibody [EPR22062] (ab222406)

    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).

     

     

  • Flow Cytometry (Intracellular) - Anti-Bcr antibody [EPR22062] (ab222406)
    Flow Cytometry (Intracellular) - Anti-Bcr antibody [EPR22062] (ab222406)

    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. 

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcr antibody [EPR22062] (ab222406)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcr antibody [EPR22062] (ab222406)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcr antibody [EPR22062] (ab222406)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcr antibody [EPR22062] (ab222406)

    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).

  • Western blot - Anti-Bcr antibody [EPR22062] (ab222406)
    Western blot - Anti-Bcr antibody [EPR22062] (ab222406)
    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).

  • Western blot - Anti-Bcr antibody [EPR22062] (ab222406)
    Western blot - Anti-Bcr antibody [EPR22062] (ab222406)
    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 minutes


    Blocking/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).

  • Anti-Bcr antibody [EPR22062] (ab222406)
    Anti-Bcr antibody [EPR22062] (ab222406)

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.

Click here to view the general protocols

References (0)

Publishing research using ab222406? Please let us know so that we can cite the reference in this datasheet.

ab222406 has not yet been referenced specifically in any publications.

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