Anti-Phospholipase C gamma 1/PLC-gamma-1 抗体 [10-PLC-gamma]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Phospholipase C gamma 1/PLC-gamma-1 antibody [10-PLC-gamma] (AB302940)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PLCG1 KO HEK293T (ab266530) cells labelling Phospholipase Cγ1 with ab302940 at 1/100 (10.63 ug/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green). Confocal image showing cytoplasmic staining in parental HEK293T cell line and no staining in PLCG1 KO HEK293T cell line is observed. ab202272 Recombinant Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab202272) was used to counterstain tubulin at 1/50 10 ug/mL dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Phospholipase C gamma 1/PLC-gamma-1 antibody [10-PLC-gamma] (AB302940)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MCF7 cells labelling Phospholipase Cγ1 with ab302940 at 1/50 (21.26 ug/ml) dilution, followed by ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green). Confocal image showing cytoplasmic staining in MCF7 cell line is observed. ab202272 Recombinant Alexa Fluor® 594 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab202272) was used to counterstain tubulin at 1/50 10 ug/mL dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.

• WB

Lab

Western blot - Anti-Phospholipase C gamma 1/PLC-gamma-1 antibody [10-PLC-gamma] (AB302940)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Lanes 1-8 : 72 seconds Lanes 9-10 : 59 seconds

All lanes:

Western blot - Anti-Phospholipase C gamma 1/PLC-gamma-1 antibody [10-PLC-gamma] (ab302940) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell), whole cell lysate at 10 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg

Lane 5:

Mouse brain tissue lysate at 10 µg

Lane 6:

Mouse heart tissue lysate at 10 µg

Lane 7:

Mouse kidney tissue lysate at 10 µg

Lane 8:

Mouse spleen tissue lysate at 10 µg

Lane 9:

Rat brain tissue lysate at 10 µg

Lane 10:

Rat heart tissue lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 150 kDa

false

• WB

Lab

Western blot - Anti-Phospholipase C gamma 1/PLC-gamma-1 antibody [10-PLC-gamma] (AB302940)

False colour image of Western blot : Anti-Phospholipase C gamma 1/PLC-gamma-1 antibody [10/PLCgamma] (ab302940 staining at 1/1000 dilution, shown in green; Rabbit anti-GAPDH antibody [16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab302940 was shown to bind specifically to Phospholipase C gamma 1/PLC-gamma-1. A band was observed at 150kDa in wild-type HEK-293 cell lysates with no signal observed at this size in the Phospholipase C gamma 1/PLC-gamma-1 knockout cell line. To generate this image, wild-type and Phospholipase C gamma 1/PLC-gamma-1 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a PVDF-FL membrane. Membranes were blocked in Odyssey diluted in equal volume of 0.1 % TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution. Performed under reducing conditions.

All lanes:

Western blot - Anti-Phospholipase C gamma 1/PLC-gamma-1 antibody [10-PLC-gamma] (ab302940) at 1/1000 dilution

Lane 1:

Wild type HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 2:

Phospholipase C gamma 1/PLC-gamma-1 knockout HEK-293T whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/10000 dilution

Lanes 1 - 4:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Observed band size: 150 kDa

false