HA500385

EIF2S1 Rabbit Polyclonal Antibody

cat.: HA500385

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, FC, IF-Cell
Clonality:	Polyclonal

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 36 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human EIF2S1 aa 116-315/315.
Positive control:	HepG2 cell lysate, A431 cell lysate, Raji cell lysate, Hela cell lysate, PC-12 cell lysate, NIH/3T3 cell lysate, mouse colon tissue lysate, rat spleen tissue lysate, human kidney tissue, Hela, A375, NCI-H441.
Subcellular location:	Stress granule.
Recommended Dilutions: WB IHC-P FC IF-Cell	1:500-1,000 1:1,900 1:500-1:1,000 1:100-1:200
Uniprot #:	SwissProt: P05198 Human \| Q6ZWX6 Mouse \| P68101 Rat
Alternative names:	EIF 2 alpha EIF 2 EIF 2A EIF 2alpha eIF-2-alpha eIF-2A EIF-2alpha EIF2 alpha EIF2 EIF2A EIF2S1 Eukaryotic translation initiation factor 2 subunit 1 alpha 35kDa Eukaryotic translation initiation factor 2 subunit 1 alpha Eukaryotic translation initiation factor 2 subunit 1 Eukaryotic translation initiation factor 2 subunit alpha IF2A_HUMAN

Images

Fig1: Western blot analysis of EIF2S1 on different lysates with Rabbit anti-EIF2S1 antibody (HA500385) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: A431 cell lysate
Lane 3: Raji cell lysate
Lane 4: Hela cell lysate
Lane 5: PC-12 cell lysate
Lane 6: NIH/3T3 cell lysate

Lysates/proteins at 10 µg/Lane.

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

Exposure time: 21 seconds;

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA500385) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of EIF2S1 on different lysates with Rabbit anti-EIF2S1 antibody (HA500385) at 1/500 dilution.

Lane 1: Mouse colon tissue lysate
Lane 2: Rat spleen tissue lysate

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 30 seconds;

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA500385) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-EIF2S1 antibody (HA500385) at 1/1,900 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA500385) at 1/1,900 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig4: Flow cytometric analysis of Hela cells labeling EIF2S1. Cells were fixed and permeabilized. Then stained with the primary antibody (HA500385, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
	Fig5: Immunocytochemistry analysis of A375 cells labeling EIF2S1 with Rabbit anti-EIF2S1 antibody (HA500385) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-EIF2S1 antibody (HA500385) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 647, HA1127) was used as the secondary antibody at 1/1,000 dilution.

Fig6: Immunocytochemistry analysis of NCI-H441 cells labeling EIF2S1 with Rabbit anti-EIF2S1 antibody (HA500385) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-EIF2S1 antibody (HA500385) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.