Anti-JMJD6 antibody [EPR26813-90] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-JMJD6 antibody [EPR26813-90] - BSA and Azide free (AB307655)

This data was developed using ab307654, the same antibody clone in a different buffer formulation. The samples were run on a Bis-Tris gel. Performed under reducing conditions. Blocking and diluting buffer and concentration : Intercept® (TBS) diluted with an equal volume of 0.1% TBS. False colour image of Western blot : Anti-JMJD6 antibody EPR26813-90 staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody 6C5 loading control staining at 1/20000 dilution, shown in red. In Western blot, ab307654 was shown to bind specifically to JMJD6. A band was observed at 50, 110-260 kDa in wild-type 293T cell lysates whereas no signal observed at this size in JMJD6 knockout cell line (ab266402). To generate this image, wild-type and JMJD6 knockout 293T cell lysates (ab257490) were analyzed. First, samples were run on a SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an 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-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) at 1/10000 dilution and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution. The molecular weight observed is consistent with what has been described in the literature (PMID : 24360279). The identity of the lower MW band at approximately 39 kDa is unknown.

All lanes:

Western blot - Anti-JMJD6 antibody [EPR26813-90] (<a href='/en-us/products/primary-antibodies/jmjd6-antibody-epr26813-90-ab307654'>ab307654</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

JMJD6 knockout 293T whole cell lysate at 20 µg

Lane 3:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 4:

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

Secondary

Lanes 1 - 4:

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

Lanes 1 - 4:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Observed band size: 50 kDa,110 kDa,260 kDa

false

• WB

Supplier Data

Western blot - Anti-JMJD6 antibody [EPR26813-90] - BSA and Azide free (AB307655)

This data was developed using ab307654, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST. Exposure time : 125 seconds.

All lanes:

Western blot - Anti-JMJD6 antibody [EPR26813-90] (<a href='/en-us/products/primary-antibodies/jmjd6-antibody-epr26813-90-ab307654'>ab307654</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 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 1/20000 dilution

Observed band size: 50 kDa,110 kDa,260 kDa

false

Exposure time: 125s