Anti-eIF2A antibody [EPR11042]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (AB169528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cerebrum tissue sections labeling eIF2A with ab169528 at 1/200 dilution (12.52 µg/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (AB169528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling eIF2A with ab169528 at 1/200 dilution (12.52 µg/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (AB169528)

Immunohistochemical analysis of paraffin embedded Human prostatic hyperplasia tissue labeling eIF2A with unpurified ab169528 antibody at 1/50.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (AB169528)

Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling eIF2A with Purified ab169528 at 1 : 100 dilution (4.3 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.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (AB169528)

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling eIF2A with purified ab169528 at 1/250 dilution. Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor^®488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (AB169528)

Immunohistochemical analysis of paraffin embedded Human pancreas tissue labeling eIF2A with unpurified ab169528 antibody at 1/50.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-eIF2A antibody [EPR11042] (AB169528)

ab169528 staining eIF2A in wild-type HAP1 cells (top panel) and EIF2A knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol for 5 minutes, permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated with ab169528 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (AB169528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cerebrum tissue sections labeling eIF2A with ab169528 at 1/200 dilution (12.52 µg/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF2A antibody [EPR11042] (AB169528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat cerebrum tissue sections labeling eIF2A with ab169528 at 1/200 dilution (12.52 µg/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

Lab

Western blot - Anti-eIF2A antibody [EPR11042] (AB169528)

All lanes:

Western blot - Anti-eIF2A antibody [EPR11042] (ab169528) at 1/1000 dilution

Lane 1:

Ramos (Human Burkitt's lymphoma Blymphocyte) whole cell lysates at 20 µg

Lane 2:

PC-12 (Rat adrenal gland pheochromocytoma) 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: 65 kDa

Observed band size: 65 kDa

false

• WB

Lab

Western blot - Anti-eIF2A antibody [EPR11042] (AB169528)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : eIF2A knockout HAP1 cell lysate (20 μg)
Lane 3 : Ramos cell lysate (20 μg)
Lane 4 : MOLT4 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab169528 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab169528 was shown to specifically react with eIF2A when eIF2A knockout samples were used. Wild-type and eIF2A knockout samples were subjected to SDS-PAGE. ab169528 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-eIF2A antibody [EPR11042] (ab169528)

Predicted band size: 65 kDa

false

• WB

Lab

Western blot - Anti-eIF2A antibody [EPR11042] (AB169528)

All lanes:

Western blot - Anti-eIF2A antibody [EPR11042] (ab169528) at 1/1000 dilution

All lanes:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates 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>) at 1/20000 dilution

Predicted band size: 65 kDa

Observed band size: 65 kDa

false