Anti-PRDM16 antibody [EPR24315-59] - BSA and Azide free

• WB

Lab

Western blot - Anti-PRDM16 antibody [EPR24315-59] - BSA and Azide free (AB303535)

This data was developed using the same antibody clone in a different buffer formulation (ab303534).

Western blot : Anti-PRDM16 antibody [EPR24315-59] (ab303534) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab303534 was shown to bind specifically to PRDM16. A band was observed at 150 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in PRDM16 knockout cell line. To generate this image, wild-type and PRDM16 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-PRDM16 antibody [EPR24315-59] (<a href='/en-us/products/primary-antibodies/prdm16-antibody-epr24315-59-ab303534'>ab303534</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

PRDM16 knockout HCT 116 cell lysate at 20 µg

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

LnCap cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 150 kDa

false

• WB

Supplier Data

Western blot - Anti-PRDM16 antibody [EPR24315-59] - BSA and Azide free (AB303535)

This data was developed using ab303535, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-PRDM16 antibody [EPR24315-59] (<a href='/en-us/products/primary-antibodies/prdm16-antibody-epr24315-59-ab303534'>ab303534</a>) at 1/1000 dilution

Lane 1:

HEK-293T cells transfected with an empty vector containing a myc-his tag, whole cell lysate at 20 µg

Lane 2:

HEK-293T cells transfected with a mouse PRDM16 expression vector containing a myc-his tag, 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 1/50000 dilution

Predicted band size: 140 kDa

Observed band size: 150 kDa,170 kDa

false

Exposure time: 1s

• WB

Supplier Data

Western blot - Anti-PRDM16 antibody [EPR24315-59] - BSA and Azide free (AB303535)

This data was developed using ab303535, the same antibody clone in a different buffer formulation.

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

Low expression :  kidney, skeletal muscle and liver (PMID : 17618855).

The molecular weight observed is consistent with what has been described in the literature (PMID : 25662275).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

Lysates were freshly made and used immediately to minimize protein degradation.

All lanes:

Western blot - Anti-PRDM16 antibody [EPR24315-59] (<a href='/en-us/products/primary-antibodies/prdm16-antibody-epr24315-59-ab303534'>ab303534</a>) at 1/1000 dilution

Lane 1:

Mouse brown fat tissue lysate at 20 µg

Lane 2:

Mouse kidney tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse liver tissue 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 1/50000 dilution

Predicted band size: 140 kDa

Observed band size: 150 kDa,170 kDa

true

Exposure time: 180s