Anti-SET7 antibody [EPR23775-142]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-SET7 antibody [EPR23775-142] (AB274416)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell, Left) / HeLa (Human cervix adenocarcinoma epithelial cell, RIght) cells labelling SET7 with ab274416 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : HepG2 cells.

• WB

Lab

Western blot - Anti-SET7 antibody [EPR23775-142] (AB274416)

Blocking and dilution buffer : Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBST

Lanes 1-3 : Merged signal (red and green). Green - ab274416 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab274416 Anti-SET7 antibody [EPR23775-142] was shown to specifically react with SET7 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265770 (knockout cell lysate ab257668) was used. Wild-type and SET7 knockout samples were subjected to SDS-PAGE. ab274416 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-SET7 antibody [EPR23775-142] (ab274416) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SET7 knockout HeLa cell lysate at 20 µg

Lane 3:

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

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 41 kDa

Observed band size: 50 kDa

false

• WB

Unknown

Western blot - Anti-SET7 antibody [EPR23775-142] (AB274416)

Blocking and dilution buffer : Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBST

Lanes 1-3 : Merged signal (red and green). Green - ab274416 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab274416 Anti-SET7 antibody [EPR23775-142] was shown to specifically react with SET7 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265770 (knockout cell lysate ab257668) was used. Wild-type and SET7 knockout samples were subjected to SDS-PAGE. ab274416 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-SET7 antibody [EPR23775-142] (ab274416) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SET7 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SETD7 (SET7) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-setd7-set7-knockout-hela-cell-line-ab265770'>ab265770</a>)

Lane 3:

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

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 41 kDa

Observed band size: 50 kDa

false

• WB

Unknown

Western blot - Anti-SET7 antibody [EPR23775-142] (AB274416)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

Negative control : HepG2 (PMID : 27183310).

All lanes:

Western blot - Anti-SET7 antibody [EPR23775-142] (ab274416) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 41 kDa

Observed band size: 50 kDa

false