EIF2S1 Recombinant antibody

EIF2S1 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

230245B8

Cat no : 82936-8-RR

Synonyms

EIF2S1/EIF2A, EIF2, EIF 2alpha, EIF 2A, eIF 2 alpha

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Validation Data Gallery

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  WB analysis using 82936-8-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 82936-8-RR (EIF2S1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  IHC staining of human ovarian cancer using 82936-8-RR

  Immunohistochemical analysis of paraffin-embedded human ovarian cancer slide using 82936-8-RR (EIF2S1 antibody) at dilution of 1:800 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 82936-8-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using EIF2S1 antibody (82936-8-RR, Clone: 230245B8 ) at dilution of 1:300 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  IF Staining of HepG2 using 82936-8-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HepG2 cells using EIF2S1 antibody (82936-8-RR, Clone: 230245B8 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-phalloidin (red).

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  FC experiment of HeLa using 82936-8-RR

  1x10^6 HeLa cells were intracellularly stained with 0.25 ug EIF2S1 Recombinant antibody (82936-8-RR, Clone:230245B8) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Isotype Control (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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  Affinity and Kinetic Characterization of 82936-8-RR

  Biolayer interferometry (BLl) kinetic assays of 82936-8-RR against Human EIF2S1 were performed. The affinity constant is below 1 pM.

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Tested Applications

• Positive WB detected in: NIH/3T3 cells, HepG2 cells, MCF-7 cells, RAW 264.7 cells
• Positive IHC detected in: human ovarian cancer; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells, HepG2 cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunohistochemistry (IHC): IHC : 1:400-1:1600
• Immunofluorescence (IF)/ICC: IF/ICC : 1:150-1:600
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

82936-8-RR targets EIF2S1 in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: EIF2S1 fusion protein Ag1645
• Full Name: eukaryotic translation initiation factor 2, subunit 1 alpha, 35kDa
• Calculated Molecular Weight: 36 kDa
• Observed Molecular Weight: 36 kDa
• GenBank Accession Number: BC002513
• Gene Symbol: EIF2S1
• Gene ID (NCBI): 1965
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

EIF2S1 is one subunit of the translation initiation factor EIF2, which catalyzes the first regulated step of protein synthesis initiation, promoting the binding of the initiator tRNA to 40S ribosomal subunits. This complex binds to a 40S ribosomal subunit, followed by mRNA binding to form a 43S preinitiation complex. Junction of the 60S ribosomal subunit to form the 80S initiation complex is preceded by hydrolysis of the GTP bound to eIF-2 and release of an eIF-2-GDP binary complex. In order for eIF-2 to recycle and catalyze another round of initiation, the GDP bound to eIF-2 must exchange with GTP by way of a reaction catalyzed by eIF-2B. EIF2A (Gene ID: 83939) and EIF2S1 (Gene ID: 1965) share the EIF2A symbol/alias in common. EIF2S1 is the alpha subunit of the eIF2 translation initiation complex. Although both of these proteins function in binding initiator tRNA to the 40S ribosomal subunit, the EIF2A protein does so in a codon-dependent manner, whereas eIF2 complex requires GTP.