Anti-KMT3A / HYPB / HIF-1 antibody [EPR20927-67] - BSA and Azide free

• WB

Lab

Western blot - Anti-KMT3A / HYPB / HIF-1 antibody [EPR20927-67] - BSA and Azide free (AB274329)

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

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, glycerol, BSA and sodium azide (ab239350).

All lanes:

Western blot - Anti-KMT3A / HYPB / HIF-1 antibody [EPR20927-67] (<a href='/en-us/products/primary-antibodies/kmt3a-hypb-hif-1-antibody-epr20927-67-ab239350'>ab239350</a>) at 1/1000 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) nuclear fraction 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 1/100000 dilution

Predicted band size: 288 kDa

Observed band size: 288 kDa

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• WB

Lab

Western blot - Anti-KMT3A / HYPB / HIF-1 antibody [EPR20927-67] - BSA and Azide free (AB274329)

False colour image of Western blot : Anti-KMT3A / HYPB / HIF-1 antibody [EPR20927-67] staining at 1/1000 dilution, shown in black; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab239350 was shown to bind specifically to KMT3A / HYPB / HIF-1. A band was observed at 120-280, 440 kDa in wild-type HeLa cell lysates with no signal observed at this size in SETD2 knockout cell line ab261792 (knockout cell lysate ab257273). To generate this image, wild-type and SETD2 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 before development with Optiblot (ECL reagent ab133456) and imaged with 20 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-KMT3A / HYPB / HIF-1 antibody [EPR20927-67] (<a href='/en-us/products/primary-antibodies/kmt3a-hypb-hif-1-antibody-epr20927-67-ab239350'>ab239350</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SETD2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SETD2 (KMT3A / HYPB / HIF-1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-setd2-kmt3a-hypb-hif-1-knockout-hela-cell-line-ab261792'>ab261792</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

Human Heart cell lysate at 20 µg

Predicted band size: 288 kDa

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• WB

Lab

Western blot - Anti-KMT3A / HYPB / HIF-1 antibody [EPR20927-67] - BSA and Azide free (AB274329)

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

Exposure times : Lane 1 : 26 seconds; Lane 2 : 86 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, glycerol, BSA and sodium azide (ab239350).

All lanes:

Western blot - Anti-KMT3A / HYPB / HIF-1 antibody [EPR20927-67] (<a href='/en-us/products/primary-antibodies/kmt3a-hypb-hif-1-antibody-epr20927-67-ab239350'>ab239350</a>) at 1/1000 dilution

Lane 1:

MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial 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 1/100000 dilution

Predicted band size: 288 kDa

Observed band size: 288 kDa

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