The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
1/1000. Detects a band of approximately 150 kDa (predicted molecular weight: 150 kDa).
Use at an assay dependent concentration. 5-10ul of antibody per 300-500 ug of total protein lysate.
Plays a role in actin reorganization and cell migration. The production of the second messenger molecules diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (IP3) is mediated by activated phosphatidylinositol-specific phospholipase C enzymes. Major substrate for heparin-binding growth factor 1 (acidic fibroblast growth factor)-activated tyrosine kinase.
The SH3 domain mediates interaction with CLNK (By similarity). The SH3 domain also mediates interaction with RALGPS1.
The receptor-mediated activation of PLC-gamma-1 and PLC-gamma-2 involves their phosphorylation by tyrosine kinases in response to ligation of a variety of growth factor receptors and immune system receptors. May be dephosphorylated by PTPRJ. Ubiquitinated by CBLB in activated T-cells.
Cell projection > lamellipodium. Cell projection > ruffle. Rapidly redistributed to ruffles and lamellipodia structures in response to epidermal growth factor (EGF) treatment.
Western blot - Anti-Phospholipase C gamma 1 antibody [M156] (ab41433)
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: Phospholipase C gamma 1 knockout HAP1 cell lysate (20 µg) Lane 3: HepG2 cell lysate (20 µg) Lane 4: Mouse brain tissue lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab41433 observed at 160 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab41433 was shown to recognize Phospholipase C gamma 1 when Phospholipase C gamma 1 knockout samples were used, along with additional cross-reactive bands. Wild-type and Phospholipase C gamma 1 knockout samples were subjected to SDS-PAGE. ab41433 diluted to 1/1000 and ab181602 (loading control to GAPDH) diluted to 1/10,000 and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
Western blot - Phospholipase C gamma 1 antibody [M156] (ab41433)
Western blot analysis of PLC gamma 1 immunoprecipitates from human jurkat cells untreated (lanes 1 & 3) or treated with pervanadate (1 mM) for 30 min (lanes 2, 4, 5 & 6). Immunoprecipitation was performed with monoclonal anti-PLC gamma 1 (ab41433). The blots were probed with anti- PLC gamma 1 (lanes 1 & 2) and anti-PLC gamma 1 (Tyr-775) (lanes 3-6). The latter antibody was used in the presence of phospho- PLC gamma 1 (Tyr-775) peptide (lane 5), or a non-specific phosphotyrosine peptide (lane 6).
Flow Cytometry-Anti-Phospholipase C gamma 1 antibody [M156](ab41433)
Overlay histogram showing Jurkat cells stained with ab41433 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab41433, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Mizrachy-Schwartz S et al. Up-regulation of AMP-activated protein kinase in cancer cell lines is mediated through c-Src activation. J Biol Chem286:15268-77 (2011).
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