ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
Use a concentration of 5 µg/ml.
1/500 - 1/2000.
Antigen retrieval with citrate buffer pH6.0 is recommended for formalin-fixed paraffin-embedded cells. For cytospin preparations of formaldehyde fixed cells permeabilization with Triton-X 100 is recommended.
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.
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).
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.
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.
Immunocytochemistry/Immunofluorescence analysis of human K562 cells labelling Bcr with ab86173 at 1/1000 (1µg/ml). An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
Western blot - Anti-Bcr antibody (ab86173)
All lanes : Anti-Bcr antibody (ab86173) at 0.1 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : 293T whole cell lysate at 50 µg
Developed using the ECL technique.
Predicted band size: 143 kDa
Exposure time: 30 seconds
Immunoprecipitation - Anti-Bcr antibody (ab86173)
Detection of Human Bcr by Immunoprecipitation, using ab86173 at 10µg/mg lysate. Image shows immunoprecipitated Bcr detected with post IP WB, loading 20% of IP and using HeLa whole cell lysate at 1mg, with an antibody recognising an upstream epitope in lane 1, ab86173 at 1 µg/ml in lane 2 and control IgG in lane 3. Detection: Chemiluminescence with an exposure time of 10 seconds.
Flow Cytometry - Anti-Bcr antibody (ab86173)
Flow Cytometric Detection of Bcr. 1 x 106 K562 cells were fixed, permeabilized, and stained with ab86173 at 0.25 µg in a 150 µl reaction. Image shows Isotype anti-KLH control (red), no antibody (blue) and ab86173 (black).
ICC/IF image of ab86173 stained HepG2 cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab86173, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.