Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (AB184845)

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.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (AB184845)

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).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (AB184845)

This data was developed using ab52607, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human astrocytoma tissue sections labeling Apolipoprotein E with purified ab52607 at 1/800 dilution (0.13 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IP

Lab

Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (AB184845)

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

All lanes:

Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] (<a href='/en-us/products/primary-antibodies/apolipoprotein-e-antibody-ep1374y-ab52607'>ab52607</a>)

Predicted band size: 36 kDa

false

• IP

Lab

Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (AB184845)

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

All lanes:

Immunoprecipitation - Anti-Apolipoprotein E antibody [EP1374Y] (<a href='/en-us/products/primary-antibodies/apolipoprotein-e-antibody-ep1374y-ab52607'>ab52607</a>)

Predicted band size: 36 kDa

false

• WB

Lab

Western blot - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (AB184845)

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.

All lanes:

Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (<a href='/en-us/products/primary-antibodies/apolipoprotein-e-antibody-ep1374y-ab52607'>ab52607</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

APOE knockout HEK293T cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 36 kDa

Observed band size: 36 kDa

false

• WB

Lab

Western blot - Anti-Apolipoprotein E antibody [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.

All lanes:

Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (<a href='/en-us/products/primary-antibodies/apolipoprotein-e-antibody-ep1374y-ab52607'>ab52607</a>) at 1/1000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

APOE knockout HepG2 cell lysate at 20 µg

Lane 2:

Western blot - Human APOE knockout Hep G2 cell line (<a href='/en-us/products/cell-lines/human-apoe-knockout-hep-g2-cell-line-ab280875'>ab280875</a>)

Lane 3:

Human Liver cell lysate at 20 µg

Lane 4:

Human Kidney cell lysate at 20 µg

Predicted band size: 36 kDa

Observed band size: 34 kDa,34-37 kDa

false

• WB

Lab

Western blot - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (AB184845)

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.

All lanes:

Western blot - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (ab184845) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TGFA knockout HeLa cell lysate at 20 µg

Lane 3:

Human liver cancer tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 36 kDa

Observed band size: 36 kDa

false

• WB

Unknown

Western blot - Anti-Apolipoprotein E antibody [EP1374Y] - Low endotoxin, Azide free (AB184845)

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

All lanes:

Western blot - Anti-Apolipoprotein E antibody [EP1374Y] (<a href='/en-us/products/primary-antibodies/apolipoprotein-e-antibody-ep1374y-ab52607'>ab52607</a>) at 1/10000 dilution

Lane 1:

Human liver lysate at 20 µg

Lane 2:

Human cerebellum lysate at 20 µg

Lane 3:

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

Lane 4:

Human serum lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 36 kDa

false