Anti-MAPK6/ERK3 antibody [EP1720Y] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAPK6/ERK3 antibody [EP1720Y] - BSA and Azide free (AB247348)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney carcinoma tissue sections labeling MAPK6/ERK3 with Purified ab53277 at 1 : 100 dilution (13.52 μg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Purified)ImmunoHistoProbe one step HRP Polymer (ready to use) was used for detection. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab53277)

• WB

Lab

Western blot - Anti-MAPK6/ERK3 antibody [EP1720Y] - BSA and Azide free (AB247348)

This data was developed using the same antibody clone in a different buffer formulation (ab53277).

Lanes 1-3 : Merged signal (red and green). Green - ab53277 observed at 90 kDa. Red - loading control ab8245 observed at 36 kDa.

ab53277 Anti-MAPK6/ERK3 antibody [EP1720Y] was shown to specifically react with 3 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264910 (knockout cell lysate ab257526) was used. Wild-type and 3 knockout samples were subjected to SDS-PAGE. ab53277 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-MAPK6/ERK3 antibody [EP1720Y] (<a href='/en-us/products/primary-antibodies/mapk6-erk3-antibody-ep1720y-ab53277'>ab53277</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

MAPK6 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human MAPK6 (ERK3) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-mapk6-erk3-knockout-hela-cell-line-ab264910'>ab264910</a>)

Lane 3:

2.4G2 (Rat B cell lymphoma B lymphocyte) cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 83 kDa

Observed band size: 90 kDa

false

• WB

Lab

Western blot - Anti-MAPK6/ERK3 antibody [EP1720Y] - BSA and Azide free (AB247348)

This data was developed using the same antibody clone in a different buffer formulation (ab53277).

Lanes 1 - 4 : Merged signal (red and green). Green - ab53277 observed at 90 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab53277 was shown to react with MAPK6 in HAP1 wild-type cells in Western blot. Loss of signal was observed when MAPK6 knockout sample was used. HAP1 wild-type and MAPK6 knockout whole cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween^®) before incubation with ab53277 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 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^® 800CW) preabsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-MAPK6/ERK3 antibody [EP1720Y] (<a href='/en-us/products/primary-antibodies/mapk6-erk3-antibody-ep1720y-ab53277'>ab53277</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

MAPK6 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

A431 whole cell lysate at 20 µg

Lane 4:

MCF7 whole cell lysate at 20 µg

Predicted band size: 83 kDa

Observed band size: 90 kDa

false