Key features and details
- Mouse monoclonal [EIF2a] to EIF2S1
- Suitable for: ICC/IF, WB, IHC-P
- Reacts with: Mouse, Rat, Human, African green monkey
- Isotype: IgG1
参阅全部 EIF2S1 一抗
描述小鼠单克隆抗体[EIF2a] to EIF2S1
经测试应用适用于: ICC/IF, WB, IHC-Pmore details
种属反应性与反应: Mouse, Rat, Human, African green monkey
Recombinant full length protein (Human).
- Human Jurkat, CEM and HeLa cells, mouse 3T3L1 and rat PC-12 cells.
存放说明Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Preservative: 0.1% Sodium azide
Concentration information loading...
纯度Protein A purified
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.
|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.|
功能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.
序列相似性Belongs to the eIF-2-alpha family.
Contains 1 S1 motif domain.
翻译后修饰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.
细胞定位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
- EIF 2 alpha antibody
- EIF 2 antibody
- EIF 2A antibody
All lanes : Anti-EIF2S1 antibody [EIF2a] (ab5369)
Lane 1 : COS-7 (african green monkey kidney fibroblast-like cell line) whole cell lysate at 20 µg
Lane 2 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 3 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 20 µg
Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 5 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 6 : NIH/3T3 (mouse embryonic fibroblast cell line) whole cell lysate
Lane 7 : A431 (human epidermoid carcinoma cell line) whole cell lysate
Lanes 1-4 & 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
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.
ab5369 被引用在 43 文献中.
- Klann K et al. Functional Translatome Proteomics Reveal Converging and Dose-Dependent Regulation by mTORC1 and eIF2a. Mol Cell 77:913-925.e4 (2020). PubMed: 31812349
- Cormerais Y et al. Inhibition of the amino-acid transporter LAT1 demonstrates anti-neoplastic activity in medulloblastoma. J Cell Mol Med 23:2711-2718 (2019). PubMed: 30784173
- Liu P et al. Crizotinib-induced immunogenic cell death in non-small cell lung cancer. Nat Commun 10:1486 (2019). PubMed: 30940805
- Ma YF et al. Phosphorylation of AKT serine/threonine kinase and abundance of milk protein synthesis gene networks in mammary tissue in response to supply of methionine in periparturient Holstein cows. J Dairy Sci 102:4264-4274 (2019). PubMed: 30879806
- Daher B et al. Genetic Ablation of the Cystine Transporter xCT in PDAC Cells Inhibits mTORC1, Growth, Survival, and Tumor Formation via Nutrient and Oxidative Stresses. Cancer Res 79:3877-3890 (2019). PubMed: 31175120