Antibodies, Proteins, Kits and Reagents for Life Science

284 784 ₸ Product size

100 µl 284 784 ₸

Order now and get it on Tuesday March 02, 2021

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: WB, IHC-P, Flow Cyt, IP
Reacts with: Mouse, Rat, Human

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 & Species

Application	Species
Flow Cyt	Human
IHC-P	Mouse Rat
IP	Human
WB	Rat Human

See all applications and species data

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: 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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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

EPR22062
Isotype

IgG
Research areas

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 Knockout 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 knockout cell line ab266583 (BCR knockout cell lysate ab257858) below 150kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and BCR knockout 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;
>C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.
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 - Anti-Bcr antibody [EPR22062] (ab222406)

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)

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)

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)

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)

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)

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