This protein is known to be similar in amino acid sequence to HCK (P08631), LCK (P06239), FYN (P06241), YES1 (P07947), and SRC (P12931). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents
This product is a recombinant rabbit monoclonal antibody.
Our Abpromise guarantee covers the use of ab40660 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
应用 | Ab评论 | 说明 |
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WB | 1/1000 - 1/10000. Detects a band of approximately 56 kDa (predicted molecular weight: 61 kDa).Can be blocked with Lyn (phospho Y396) peptide (ab202645). Induction of cells with Pervanadate: Prepare 500mM stock Sodium Orthovanadate in PBS. To make 100mM (100x) Pervanadate Na3VO4 right before experiment, add 400ul 500mM Sodium Orthovanadate to 1560ul H2O and then 40ul H2O2, vortex and incubate at room RT for 5-10min. Cells are serum-starved overnight, and then treated with freshly made Pervanadate (100x, final concentration 1mM) for 20min at 37°C. |
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Flow Cyt | 1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF | Use at an assay dependent concentration. |
Blocking and dilution buffer: 5% BSA/TBST.
Blocking and dilution buffer: 5% BSA/TBST.
Blocking and dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/Immunofluorescence analysis of HeLa cells (untreated and treated with 1mM pervanadate for 15 minutes) labelling SRC Family (phospho Y418) with purified ab40660 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/250) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
ICC/IF image of unpurified ab40660 stained PC12 cells. The cells were 4% formaldehyde fixed (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 (ab40660, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.
Flow Cytometry analysis of HeLa cells (treated with 1mM pervanadate for 30 minutes) labelling SRC Family (phospho Y418) with purified ab40660 at 1/80 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody.
Black - Isotype control, rabbit monoclonal IgG with pervanade treated HeLa cells.
Blue - Unlabelled control, pervanadate treated HeLa cells without incubation with primary and secondary antibodies.
Red - Pervandate treated HeLa cells with ab40660.
Green - Untreated HeLa cells labelled with ab40660.
Direct ELISA antibody dose-response curve using unpurified ab40660 at 0-1000 ng/ml. Antigen concentration of 1000 ng/mL. An alkaline phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"