MA1-153

antibody from Invitrogen Antibodies

Targeting: BCR ALL, BCR1, CML, D22S11, D22S662, PHL

Western blot

Immunoprecipitation

Immunohistochemistry

Antibody data

Submit

Validation data

Product number: MA1-153 - Provider product page
Provider: Invitrogen Antibodies
Product name: BCR-ABL Monoclonal Antibody (7C6)
Antibody type: Monoclonal
Antigen: Other
Description: 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.
Reactivity: Human, Mouse
Host: Mouse
Isotype: IgG
Antibody clone number: 7C6
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: -20°C

BCR protein structure - MA1-153 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of BCR-ABL (green) in K562 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a BCR-ABL monoclonal antibody (Product # MA1-153) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502). Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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 4øC. 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 4øC, 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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2. The TF1-BAP model displays similar features as chronic myeloid leukemia (CML) persistent cells. (A) Strategy for generating TF1-BA and TF1-BAP cell lines. (B) Representative images of BMPR1B staining of TF1-BA and TF1-BAP cells. (C) Western blot analysis showing BMPR1B levels in TF1-BA and TF1-BAP cells; Scatter plot showing BMPR1B (GAPDH used as an internal control) n=5. (D) Representative FACS plots of TF1-BA and TF1-BAP cells analyzed for their CD34 and CD38 content. (E) Dot plot showing colony forming cell (CFC) activity from TF1-BA and TF1-BAP cells. (F) Dot plot showing long-term culture-initiating cell (LTC-IC) activity from TF1-BA and TF1-BAP cells. (G) Western blot analysis showing BCR-Abl and P-CRKL levels in TF1-BA and TF1-BAP cells. Bar graph showing BCR-Abl and P-CRKL (GAPDH used as an internal control); n=4. (H) TF1-BA and TF1-BAP cells were analyzed for their content in P-Smad1/5/8 and P-Stat3. Dot plots represent percentage of PSmad1/ 5/8 (left panel), P-Stat3 (middle panel) or double positive P-Smad1/5/8-P-Stat3 (right panel). (E-H) Unpaired t -test significant: * P