Anti-EIF2S1 antibody (ab26197)

Western blot - eIF2 alpha antibody (ab26197)

All lanes : Anti-EIF2S1 antibody (ab26197) at 1 µg/ml

Lane 1 : HeLa whole cell lysate
Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)

Lysates/proteins at 20 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 36 kDa
Observed band size : 36 kDa

ab26197 detects a band of ~ 36 kDa in HeLa, Jurkat and A431 whole cell lysates. This corresponds to the predicted molecular weight for eIF2 alpha according to Swiss prot. A number of other bands are detected at ~ 48, 23 and 15 kDa. We do not know the identity of these bands and we predict they represent non-specific binding.

Western blot - eIF2 alpha antibody (ab26197)

All lanes : Anti-EIF2S1 antibody (ab26197) at 1 µg/ml

Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Performed under reducing conditions.

Predicted band size : 36 kDa
Observed band size : 36 kDa

Immunocytochemistry/ Immunofluorescence - eIF2 alpha antibody (ab26197)

ICC/IF image of ab26197 stained human HeLa cells. The cells were 4% PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab26197, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - eIF2 alpha antibody (ab26197)

Image courtesy of Human Protein Atlas

ab26197 staining eiF2 alpha in human small intestine. The paraffin embedded tissue was incubated with ab26197 (1/40 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6.

ab26197 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

Further results for this antibody can be found at www.proteinatlas.org