Anti-Apolipoprotein E 抗体 [EP1374Y] - Low endotoxin, Azide free (ab184845)

False colour image of Western blot: Anti-Apolipoprotein E antibody [EP1374Y] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab52607 was shown to bind specifically to Apolipoprotein E. A band was observed at 34-37 kDa in wild-type HepG2 cell lysates with no signal observed at this size in APOE knockout cell line. To generate this image, wild-type and APOE knockout HepG2 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 (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

This data was developed using ab52607, the same antibody clone in a different buffer formulation.

This data was developed using ab52607, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Apolipoprotein E with Purified ab52607 at 1:50 dilution (2.0 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation (ab52607).

Lanes 1-3: Merged signal (red and green). Green - ab52607 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa. 

 ab52607 Anti-Apolipoprotein E antibody [EP1374Y] was shown to specifically react with Apolipoprotein E in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266546 (knockout cell lysate ab256838) was used.  Wild-type and Apolipoprotein E knockout samples were subjected to SDS-PAGE.  ab52607 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively.  Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation (ab52607).

Lanes 1-3: Merged signal (red and green). Green - ab52607 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa.

ab52607 Anti-Apolipoprotein E antibody [EP1374Y] was shown to specifically react with Apolipoprotein E in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265166 (knockout cell lysate ab257739) was used. Wild-type and Apolipoprotein E knockout samples were subjected to SDS-PAGE. ab52607 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling Apolipoprotein E (red) with purified ab52607 at a 1/250 dilution (10ug/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti rabbit IgG (Alexa Fluor^®488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue). 

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52607). 

This data was developed using ab52607, the same antibody clone in a different buffer formulation.
Purified ab52607 at 1/20 dilution (0.5µg) immunoprecipitating Apolipoprotein E in HepG2 whole cell lysate.
Lane 1 (input): HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab52607 + HepG2 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52607 in HepG2 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 36 kDa

This data was developed using ab52607, the same antibody clone in a different buffer formulation.
Purified ab52607 at 1/20 dilution (0.5µg) immunoprecipitating Apolipoprotein E in Human serum.
Lane 1 (input): Human serum 10µg
Lane 2 (+): ab52607 + Human serum.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52607 in Human serum.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 36 kDa