Anti-LDL Receptor antibody [EP1553Y]

• WB

Lab

Western blot - Anti-LDL Receptor antibody [EP1553Y] (AB52818)

Anti-LDLR antibody [EP1553Y] (ab52818) 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, ab52818 was shown to bind specifically to LDLR. A band was observed at 130/160 kDa in wild-type HeLa cell lysates with no signal observed at this size in LDLR knockout cell line ab273838 (knockout cell lysate ab273792). To generate this image, wild-type and LDLR 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 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 [EP1553Y] (ab52818) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

LDLR knockout HeLa cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Lane 3:

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: 130 kDa,160 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LDL Receptor antibody [EP1553Y] (AB52818)

Immunohistochemical analysis of paraffin-embedded human liver sections labeling LDL Receptor with purified ab52818 at dilution of 1 : 500. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LDL Receptor antibody [EP1553Y] (AB52818)

Immunohistochemical analysis of paraffin-embedded human hepatocellular carcinoma tissue labeling LDL Receptor with ab52818 at 1/100 dilution followed by goat anti-rabbit IgG H&L (HRP) (ab97051 1/500). The sample was counterstained with hematoxylin.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LDL Receptor antibody [EP1553Y] (AB52818)

Immunohistochemical analysis of formalin fixed paraffin embedded human liver labelling LDL Receptor with ab52818 at a concentration of 0.5μg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab52818 anti-LDL Receptor antibody [EP1553Y] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• WB

Unknown

Western blot - Anti-LDL Receptor antibody [EP1553Y] (AB52818)

LDL Receptor contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

All lanes:

Western blot - Anti-LDL Receptor antibody [EP1553Y] (ab52818) at 1/500 dilution

All lanes:

Human plasma total protein lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 95 kDa

Observed band size: 100 kDa,27 kDa,48 kDa

true

Exposure time: 4min

• WB

Unknown

Western blot - Anti-LDL Receptor antibody [EP1553Y] (AB52818)

Blocking/Diluting buffer 5% NFDM/TBST

All lanes:

Western blot - Anti-LDL Receptor antibody [EP1553Y] (ab52818) at 1/5000 dilution

All lanes:

Mouse liver at 15 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>)

Predicted band size: 95 kDa

Observed band size: 140 kDa

false

• WB

Lab

Western blot - Anti-LDL Receptor antibody [EP1553Y] (AB52818)

Blocking buffer : 5% NFDM/TBST

Diltuion buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-LDL Receptor antibody [EP1553Y] (ab52818) at 1/1000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg

Lane 2:

HeLa treated with GW3965 for 8 hours at the final concentration of 5uM whole cell lysates at 20 µg

Lane 3:

HeLa treated with GW3965 for 24 hours at the final concentration of 5uM whole cell lysates at 20 µg

Lane 4:

Raw264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysates at 20 µg

Lane 5:

Raw264.7 treated with GW3965 for 8 hours at the final concentration of 5uM whole cell lysates at 20 µg

Lane 6:

Raw264.7 treated with GW3965 for 24 hours at the final concentration of 5uM whole cell lysates 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

Predicted band size: 95 kDa

Observed band size: 140 kDa

false

Exposure time: 1min

• WB

Lab

Western blot - Anti-LDL Receptor antibody [EP1553Y] (AB52818)

Blocking buffer : 5% NFDM/TBST

Diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-LDL Receptor antibody [EP1553Y] (ab52818) at 1/1000 dilution

Lane 1:

PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

LNCaP (Human prostate 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:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

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

Lane 6:

A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

Predicted band size: 95 kDa

Observed band size: 150 kDa

false

• WB

Lab

Western blot - Anti-LDL Receptor antibody [EP1553Y] (AB52818)

Blocking and diluting buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with the literature (PMID : 15199428, PMID : 8349823, PMID : 10906332, PMID : 24918045).

All lanes:

Western blot - Anti-LDL Receptor antibody [EP1553Y] (ab52818) at 0.4 µg/mL

All lanes:

HepG2 (Human hepatocellular carcinoma 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

Predicted band size: 95 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-LDL Receptor antibody [EP1553Y] (AB52818)

Blocking buffer : 5% NFDM/TBST

Diltuion buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-LDL Receptor antibody [EP1553Y] (ab52818) at 1/1000 dilution

Lane 1:

Mouse liver lysate at 20 µg

Lane 2:

Mouse lung lysate at 20 µg

Lane 3:

Mouse colon lysate at 20 µg

Lane 4:

Human liver lysate at 20 µg

Lane 5:

HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate at 20 µg

Lane 6:

HeLa (Human epithelial cell line from cervix adenocarcinoma) 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 1/1000 dilution

Predicted band size: 95 kDa

Observed band size: 140 kDa

false

Exposure time: 3min