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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
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 47 文献中.
- van Ziel AM et al. No evidence for cell-to-cell transmission of the unfolded protein response in cell culture. J Neurochem 152:208-220 (2020). PubMed: 31442299
- 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
- Costa R et al. Impaired Mitochondrial ATP Production Downregulates Wnt Signaling via ER Stress Induction. Cell Rep 28:1949-1960.e6 (2019). PubMed: 31433973
- Chiritoiu M et al. GRASP55 and UPR Control Interleukin-1ß Aggregation and Secretion. Dev Cell 49:145-155.e4 (2019). PubMed: 30880003
- Ebrahimi SM et al. L-lysine protects C2C12 myotubes and 3T3-L1 adipocytes against high glucose damages and stresses. PLoS One 14:e0225912 (2019). PubMed: 31856203