製品の概要

  • 製品名
    Anti-EIF2S1 antibody [EIF2a]
    EIF2S1 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [EIF2a] to EIF2S1
  • アプリケーション
    適用あり: ICC/IF, WB, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Recombinant full length protein (Human).

  • エピトープ
    not mapped.
  • ポジティブ・コントロール
    • Human Jurkat, CEM and HeLa cells, mouse 3T3L1 and rat PC-12 cells.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab5369 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 5 µg/ml.
WB 1/500 - 1/1000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
IHC-P Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ターゲット情報

  • 機能
    Functions in the early steps of protein synthesis by forming a ternary complex with GTP and initiator tRNA. 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.
  • 配列類似性
    Belongs to the eIF-2-alpha family.
    Contains 1 S1 motif domain.
  • 翻訳後修飾
    Substrate for at least 4 kinases: EIF2AK1/HRI, EIF2AK2/PKR, EIF2AK3/PERK and EIF2AK4/GCN2. Phosphorylation stabilizes the eIF-2/GDP/eIF-2B complex and prevents GDP/GTP exchange reaction, thus impairing the recycling of eIF-2 between successive rounds of initiation and leading to global inhibition of translation (PubMed:15207627, PubMed:18032499). Phosphorylated; phosphorylation on Ser-52 by the EIF2AK4/GCN2 protein kinase occurs in response to amino acid starvation and UV irradiation.
  • 細胞内局在
    Cytoplasmic granule. The cytoplasmic granules are stress granules which are a dense aggregation in the cytosol composed of proteins and RNAs that appear when the cell is under stress. Colocalizes with NANOS3 in the stress granules (By similarity).
  • Information by UniProt
  • 参照データベース
  • 別名
    • EIF 2 alpha antibody
    • EIF 2 antibody
    • EIF 2A antibody
    • EIF 2alpha antibody
    • eIF-2-alpha antibody
    • eIF-2A antibody
    • EIF-2alpha antibody
    • EIF2 alpha antibody
    • EIF2 antibody
    • EIF2A antibody
    • EIF2S1 antibody
    • Eukaryotic translation initiation factor 2 subunit 1 alpha 35kDa antibody
    • Eukaryotic translation initiation factor 2 subunit 1 alpha antibody
    • Eukaryotic translation initiation factor 2 subunit 1 antibody
    • Eukaryotic translation initiation factor 2 subunit alpha antibody
    • IF2A_HUMAN antibody
    see all

Anti-EIF2S1 antibody [EIF2a] 画像

  • All lanes : Anti-EIF2S1 antibody [EIF2a] (ab5369)

    Lane 1 : COS-7 whole cell extracts at 20 µg
    Lane 2 : MCF7 whole cell extracts at 20 µg
    Lane 3 : PC-12 at 20 µg
    Lane 4 : HeLa lysate
    Lane 5 : Jurkat
    Lane 6 : NIH/3T3
    Lane 7 : A-431

    Secondary
    Lanes 1, 2, 3, 4, 6 & 7 : Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate at 1/4000 dilution
    Lane 5 : Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate at 4000


    Predicted band size : 36 kDa
  • Immunohistochemical analysis of human colon carcinoma (right) compared to a negative control (left) using ab5369 at the dilution 1/10.

  • ICC/IF image of ab5369 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab5369, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Ab5369 staining EIF2S1 in human placenta. Staining is localised to the cytoplasm.
    Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

Anti-EIF2S1 antibody [EIF2a] (ab5369) 使用論文

This product has been referenced in:
  • Yan L  et al. Icariside II Effectively Reduces Spatial Learning and Memory Impairments in Alzheimer's Disease Model Mice Targeting Beta-Amyloid Production. Front Pharmacol 8:106 (2017). Read more (PubMed: 28337142) »
  • Plate L  et al. Small molecule proteostasis regulators that reprogram the ER to reduce extracellular protein aggregation. Elife 5:N/A (2016). WB ; Human . Read more (PubMed: 27435961) »

See all 25 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Loading amount
60 µg
Specification
Liver
Gel Running Conditions
Reduced Denaturing (gel 10%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Jan 24 2012

Thank you for your enquiry. I am sorry it has taken some time to confirm this information with the originator for this antibody. I am sorry to confirm that the only cross reactivity data we have for ab5369 Anti-EIF2S1 antibody [EIF2a] is that it...

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Cell lysate - whole cell (S2R+ cells)
Loading amount
25 µg
Specification
S2R+ cells
Gel Running Conditions
Reduced Denaturing (4-20% tris-glycine)
Blocking step
normal horse serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Natalie Farny

Verified customer

投稿 Oct 22 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa cell line)
Loading amount
20 µg
Specification
HeLa cell line
Gel Running Conditions
Reduced Denaturing (12% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Mirjam Eckert

Verified customer

投稿 Apr 02 2009

Thank you for your enquiry. I can confirm that both these eIF2a antibodies (ab1126, ab5369)will detect both phosphorylated and de-phosphorylated forms of the protein. I hope this information is helpful. Should you have any further questions, pl...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (RAW 264.7 murine macrophage)
Loading amount
10 µg
Specification
RAW 264.7 murine macrophage
Blocking step
Other as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

投稿 Oct 23 2006

Thank you for this information and clarification. I have changed the online datasheet to reflect the fact that the antibody is raised against eIF2 alpha. Thank you again for your assistance.

Thank you for your enquiry. Our designation comes directly from Swiss-Prot where the synonyms for the protein are as follows: Eukaryotic translation initiation factor 2 alpha subunit eIF-2-alpha EIF-2alpha EIF-2A I believe the datasheet is...

Read More

We have received the following information from the source of this antibody: "The Mouse (monoclonal) anti-eIF2alpha antibody ab5369 clone EIF2alpha was made using recombinant protein. There is unfortunately no information on the epitope or region th...

Read More

I enquired with the supplier of ab5369 and they were able to provide me with the following information: approximately 30-40 micrograms of lysate was loaded. If you have any more questions, do not hesitate to contact us again.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

登録