重组Anti-IQGAP1抗体[EPR5220]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IQGAP1 antibody [EPR5220] (AB133490)

Immunohistochemical analysis of IQGAP1 in paraffin embedded Human kidney tissue using ab133490 at a dilution of 1/100.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-IQGAP1 antibody [EPR5220] (AB133490)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling IQGAP1 with unpurified ab133490 at 1/20 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-IQGAP1 antibody [EPR5220] (AB133490)

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling IQGAP1 with Purified ab133490 at 1/500 dilution (5 μg/ml). Cells were fixed with 100% methanol. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

• WB

Lab

Western blot - Anti-IQGAP1 antibody [EPR5220] (AB133490)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : IQGAP knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : HEK293 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab133490 observed at 190 kDa. Red - loading control, ab7291, observed at 52 kDa.
ab133490 was shown to specifically react with IQGAP1 when IQGAP1 knockout samples were used. Wild-type and IQGAP1 knockout samples were subjected to SDS-PAGE. ab133490 and ab7291 (loading control to alpha tubulin) were diluted 1/1000 and 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-IQGAP1 antibody [EPR5220] (ab133490)

Predicted band size: 189 kDa

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• WB

Unknown

Western blot - Anti-IQGAP1 antibody [EPR5220] (AB133490)

All lanes:

Western blot - Anti-IQGAP1 antibody [EPR5220] (ab133490) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

293T cell lysate at 10 µg

Lane 3:

Human placenta lysate at 10 µg

Lane 4:

Mouse thymus lysate at 10 µg

Lane 5:

F9 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 189 kDa

Observed band size: 195 kDa

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