Invitrogen

BCR-ABL Monoclonal Antibody (7C6)

Advanced Verification

This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated.

1 Published Figure

1 Reference

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Datasheet

Protocols

Questions & Answers

Tech Support

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BCR-ABL Antibody in Western Blot (WB) — Knockout of BCR was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR981967_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of BCR was performed by loading 30 µg of A-431 Wild Type (Lane 1), A-431 Cas9 (Lane 2) andA-431 BCR KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-BCR-ABL Monoclonal Antibody (7C6) (Product # MA1-153, 1:1,000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to BCR.

BCR-ABL Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of BCR-ABL was performed using 70% confluent log phase K-562 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with BCR-ABL Monoclonal Antibody (7C6) (Product # MA1-153) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing membranous localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

BCR-ABL Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of BCR-ABL showing staining in the nucleus and cytoplasm of paraffin-embedded mouse liver tissue (right) compared with a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a BCR-ABL monoclonal antibody (Product # MA1-153) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

BCR-ABL Antibody in Immunoprecipitation (IP) — Immunoprecipitation of BCR-ABL and BCR was performed using K562 whole cell lysate. Antigen-antibody complexes were formed by incubating 800 µg of lysate with 5 µg of a BCR monoclonal antibody (Product # MA1-153) overnight on a rocking platform at 4°C. The immune complexes were captured on 50 µl Protein A/G Agarose (Product # 20421), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). The sample was resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween for 1 hour. The membrane was probed with a phosphotyrosine monoclonal antibody (detecting p-Tyr-BCR-ABL/p-Tyr-BCR) at a dilution of 1:1000 overnight rotating at 4°C, washed in TBST, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).

BCR-ABL Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A loss of signal was observed for target protein inBCR KO cell line compared to control cell line using Anti-BCR-ABL Monoclonal Antibody (7C6) (Product # MA1-153). {KO}

Product Details

MA1-153

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:1,000	View 1 publication 1 publication
Immunohistochemistry (Paraffin) (IHC (P))	1:50-1:500	-
Immunocytochemistry (ICC/IF)	1:50-1:100	-
Immunoprecipitation (IP)	5 µg	-

Product Specifications
Species Reactivity	Human, Mouse
Published species	Human
Host/Isotype	Mouse / IgG2a
Class	Monoclonal
Type	Antibody
Clone	7C6
Immunogen	Bcr686 thyroglobulin conjugate corresponding to human BCR sequence 686-696 (SSINEEITPRRQS) View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Protein A
Storage buffer	PBS with 1mg/mL BSA
Contains	0.05% sodium azide
Storage conditions	-20°C
RRID	AB_2536861

Product Specific Information

Western blot analysis of MA1-153 detects ~130-160 kDa and a 210-250 kDa protein in K562 cells. In addition to BCR-ABL, BCR and ABL are also expressed in these cells and full length BCR is also detected by the antibody. For detection purposes, non-reducing conditions resulted in enhanced detection of BCR-ABL.

Target Information

A reciprocal translocation between chromosomes 22 and 9 produces the BCR-ABL fusion oncoprotein, which is found in patients with chronic myelogenous leukemia (CML). Although there are various versions, p210 is the most commonly found BCR-ABL form. The fusion to BCR, results in deregulated/constitutively active tyrosine kinase activity of BCR-ABL which is sufficient of oncogenic transformation of myeloid cells. Inhibition of BCR-ABL oncogenic activity by the ABL-kinase inhibitor STI-571 (also known as gleevec, imatinib) became the first and best example for successful targeted therapy of cancer, and specifically CML. In recent years, several mutations (such as the T351I mutation) were found in CML patients that resulted in STI-571 resistance and the need for additional drug development.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

The quiescent fraction of chronic myeloid leukemic stem cells depends on BMPR1B, Stat3 and BMP4-niche signals to persist in patients in remission.

Haematologica

Jeanpierre S,Arizkane K,Thongjuea S,Grockowiak E,Geistlich K,Barral L,Voeltzel T,Guillemin A,Gonin-Giraud S,Gandrillon O,Nicolini FE,Mead AJ,Maguer-Satta V,Lefort S

MA1-153 was used in Western Blotting to study the involvement of bone morphogenetic protein signaling in favoring residual leukemic stem cell persistence in the bone marrow of patients having achieved remission under treatment.

Fri Jan 01 00:00:00 EST 2021

The quiescent fraction of chronic myeloid leukemic stem cells depends on BMPR1B, Stat3 and BMP4-niche signals to persist in patients in remission.

Haematologica

Jeanpierre S,Arizkane K,Thongjuea S,Grockowiak E,Geistlich K,Barral L,Voeltzel T,Guillemin A,Gonin-Giraud S,Gandrillon O,Nicolini FE,Mead AJ,Maguer-Satta V,Lefort S

Fri Jan 01 00:00:00 EST 2021

Bioinformatics

Protein Aliases: BCR/FGFR1 chimera protein; breakpoint cluster region; breakpoint cluster region homolog; Breakpoint cluster region protein; FGFR1/BCR chimera protein; Renal carcinoma antigen NY-REN-26

Gene Aliases: 5133400C09Rik; AI561783; AI853148; ALL; BCR; BCR1; CML; D22S11; D22S662; Kiaa3017; mKIAA3017; PHL

UniProt ID: (Human) P11274, (Mouse) Q6PAJ1

Entrez Gene ID: (Human) 613, (Mouse) 110279

Molecular Function: G-protein modulator