Anti-GNAQ antibody [EPR20978]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GNAQ antibody [EPR20978] (AB210004)

Immunohistochemical analysis of paraffin-embedded human mammary gland tissue labeling GNAQ with ab210004 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining on human mammary gland is observed (PMID : 22421440; PMID : 1946421; PMID : 18799799). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GNAQ antibody [EPR20978] (AB210004)

Immunohistochemical analysis of paraffin-embedded human papillary thyroid carcinoma tissue labeling GNAQ with ab210004 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining on human mammary gland is observed (PMID : 22421440; PMID : 1946421; PMID : 18799799). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Unknown

Immunoprecipitation - Anti-GNAQ antibody [EPR20978] (AB210004)

GNAQ was immunoprecipitated from 0.35 mg of A549 (human lung carcinoma cell line) whole cell lysate with ab210004 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab210004 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : A549 whole cell lysate 10 μg (Input).

Lane 2 : ab210004 IP in A549 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab210004 in A549 whole cell lysate.

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

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-GNAQ antibody [EPR20978] (ab210004)

Predicted band size: 42 kDa

Observed band size: 42 kDa

true

• WB

Lab

Western blot - Anti-GNAQ antibody [EPR20978] (AB210004)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : GNAQ knockout HAP1 whole cell lysate (20 μg)

Lanes 1 - 2 : Merged signal (red and green). Green - ab210004 observed at 42 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab210004 was shown to specifically react with GNAQ in wild-type HAP1 cells as signal was lost in GNAQ knockout cells. Wild-type and GNAQ knockout samples were subjected to SDS-PAGE. ab210004 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

Lane 1:

Western blot - Anti-GNAQ antibody [EPR20978] (ab210004)

Lane 2:

Western blot - Anti-GNAQ antibody [EPR20978] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/gnaq-antibody-epr20978-bsa-and-azide-free-ab225744'>ab225744</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

GNAQ knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 42 kDa

false

• WB

Supplier Data

Western blot - Anti-GNAQ antibody [EPR20978] (AB210004)

ab210004 was shown to react with GNAQ in wild-type HAP1 cells in Western blot with loss of signal observed in a GNAQ knockout cell line. Wild-type HAP1 and GNAQ knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab210004 overnight at 4°C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-GNAQ antibody [EPR20978] (ab210004) at 1/1000 dilution

Lane 1:

Wild-type HAP1 lysate at 30 µg

Lane 2:

GNAQ knock-out HAP1 lysate at 30 µg

Observed band size: 42 kDa

false

• WB

Supplier Data

Western blot - Anti-GNAQ antibody [EPR20978] (AB210004)

Exposure time : Lanes 1-3 & 5-7 : 3 minutes; Lane 4 : 3 seconds.

Blocking and dilution buffer : 5% NFDM/TBST

Lanes 1 - 4:

Western blot - Anti-GNAQ antibody [EPR20978] (ab210004) at 1/2000 dilution

Lanes 5 - 7:

Western blot - Anti-GNAQ antibody [EPR20978] (ab210004) at 1/1000 dilution

Lane 1:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

A549 (human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 5:

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

Lane 6:

RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 7:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

true

• WB

Supplier Data

Western blot - Anti-GNAQ antibody [EPR20978] (AB210004)

Exposure time : Lanes 2-4 : 3 minutes; Lanes 1 & 5 : 15 seconds.

Blocking and dilution buffer : 5% NFDM/TBST

Lanes 1 - 4:

Western blot - Anti-GNAQ antibody [EPR20978] (ab210004) at 1/2000 dilution

Lane 5:

Western blot - Anti-GNAQ antibody [EPR20978] (ab210004) at 1/1000 dilution

Lane 1:

Human fetal pancreas lysate at 20 µg

Lane 2:

Mouse lung lysate at 20 µg

Lane 3:

Mouse kidney lysate at 20 µg

Lane 4:

Rat lung lysate at 20 µg

Lane 5:

Human stomach lysate at 10 µg

Secondary

Lanes 1 - 4:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Lane 5:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/4000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

true

• WB

Supplier Data

Western blot - Anti-GNAQ antibody [EPR20978] (AB210004)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-GNAQ antibody [EPR20978] (ab210004) at 1/1000 dilution

Lane 1:

His-tagged human GNAQ (aa1-359) recombinant protein at 0.01 µg

Lane 2:

His-tagged human GNA11 (aa1-359) recombinant protein at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

true

Exposure time: 1s