Anti-LDL Receptor antibody [EPR24874-56]

• WB

Lab

Western blot - Anti-LDL Receptor antibody [EPR24874-56] (AB271189)

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

LDLR is a glycosylated protein and can be deglycosylated by Protein Deglycosylation MIX II.

This blot was developed using a higher sensitivity ECL substrate.

Exposure time : 92 seconds

All lanes:

Western blot - Anti-LDL Receptor antibody [EPR24874-56] (ab271189) at 1/1000 dilution

Lane 1:

Untreated Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

Huh7 whole cell lysate treated with Protein Deglycosylation MIX II at 15 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: 95 kDa

Observed band size: 120 kDa,160 kDa

false

• WB

Lab

Western blot - Anti-LDL Receptor antibody [EPR24874-56] (AB271189)

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

The expression profiles/observed MW are consistent with what has been described in the literature (PMID : 3005267).

Low expression : HEK-293 (PMID : 30617148).

Exposure time : 70 seconds

All lanes:

Western blot - Anti-LDL Receptor antibody [EPR24874-56] (ab271189) at 1/1000 dilution

Lane 1:

PC-3 (human prostate 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:

Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HEK-293 (human embryonic kidney 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: 95 kDa

Observed band size: 120 kDa,160 kDa

false

• WB

Lab

Western blot - Anti-LDL Receptor antibody [EPR24874-56] (AB271189)

Anti-LDLR antibody [EPR24874-56] (ab271189) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab271189 was shown to bind specifically to LDLR. A band was observed at 120, 150 kDa in wild-type HeLa cell lysates with no signal observed at this size in LDLR CRISPR-Cas9 edited cell line ab273838 (CRISPR-Cas9 edited cell lysate ab273792). The band observed in the CRISPR-Cas9 edited lysate lane below 120, 150 kDa is likely to represent a possible truncated form of LDLR. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and LDLR CRISPR-Cas9 edited 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 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-LDL Receptor antibody [EPR24874-56] (ab271189) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human LDLR (LDL Receptor) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ldlr-ldl-receptor-knockout-hela-cell-line-ab273838'>ab273838</a>)

Lane 2:

LDLR CRISPR-Cas9 edited HeLa cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

A431 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

Predicted band size: 95 kDa

Observed band size: 120 kDa,150 kDa

false