Anti-LDL Receptor antibody [EP1553Y] (ab52818)

Blocking buffer: 5% NFDM/TBST

Diltuion buffer: 5% NFDM/TBST

Blocking and diluting buffer: 5% NFDM/TBST.

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

Blocking buffer: 5% NFDM/TBST

Diltuion buffer: 5% NFDM/TBST

Overlay histogram showing 4% paraformaldehyde fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling LDL Receptor (red) with purified ab52818 at dilution of 1/70. The secondary antibody used was an Alexa Fluor^® 488 goat-anti-rabbit IgG at dilution of 1/2000. A non-specific IgG antibody (rabbit monoclonal) was used as isotype control (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

Overlay histogram showing U937 (Human histiocytic lymphoma cell line) cells stained with ab52818 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52818, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit monoclonal IgG (1µg/1x10⁶ cells) used under the same conditions.

Acquisition of >5,000 events was performed.

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling LDL Receptor with purified ab52818 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized using 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor^®594-conjugated goat anti-mouse IgG (1/1000) as the secondary. Nuclei were counterstained with DAPI (blue).

For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (ab150120) were used.

For negative control 2, mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) were used. 

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.

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.