Anti-eIF-2α Antibody, clone 3M6 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1560 - recombinant, expressed in HEK 293 cells
Synonym: Eukaryotic translation initiation factor 2 subunit 1
Product Type: Chemical
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:1,000 dilution of Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:1,000 dilution of Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:1,000 dilution of Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HUVEC was performed using a 1:1,000 dilution of Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytosol
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HUVEC was performed using a 1:1,000 dilution of Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytosol
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HUVEC was performed using a 1:1,000 dilution of Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytosol
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cells was performed using a 1:1,000 dilution of Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Cytosol.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from HeLa cells (Lane 1) and Raji cells (Lane 2) were probed with Cat. No. ZRB1560, Anti-eIF-2a , clone 3M6 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates eIF-2a (~36 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 10 mg/mL of non-specific GST protein (Run 1) or recombinant eIF-2a protein (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1560, Anti-eIF-2α, clone 3M6 ZooMAb® Rabbit Monoclonal. The binding of this antibody to eIF-2a protein displayed a KD of 2.6 x 10-8, whereas binding to non-specific GST protein displayed a KD of 1.5 x 10-3.
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million Ramos cells was performed using 0.1 μg of Cat. No. ZRB1560, Anti-eIF-2a, clone 3M6 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (Grey histogram), followed by a PE-conjugated Donkey Anti-Rabbit IgG secondary antibody.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 3M6, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | Unknown |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 36.11 kDa |
| observed mol wt ~36 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_004085  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| flow cytometry: suitable | |
| immunocytochemistry: suitable | |
| western blot: suitable | |
| UniProt accession no. | P05198 |
| Application: | Quality Control Testing Evaluated by Western Blotting in HeLa cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected eIF-2a in HeLa cell lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected eIF-2a in Raji cell lysate. Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected eIF-2a in HeLa cell line, A431, HUVEC, and NIH 3T3 cells. Affinity Binding Assay:: A representative lot of this antibody bound eIF-2a with a KD of 2.6 x 10-8 in an affinity binding assay. Flow Cytometry Analysis: 0.1 μg from a representative lot detected eIF-2a in one million Ramos cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them. |
| Immunogen: | His-tagged recombinant full-length human Eukaryotic translation initiation factor 2 subunit a (eIF-2a ) |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 3M6 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects Eukaryotic translation initiation factor 2a (eIF-2a). |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Eukaryotic translation initiation factor 2 subunit 1 (UniProt: P05198; also known as Eukaryotic translation initiation factor 2 subunit alpha, eIF-2-alpha, eIF-2A, eIF-2alpha) is encoded by the EIF2S1 (also known as EIF2A) gene (Gene ID: 1965) in human. eIF-2A is a heterotrimer composed of a, b, and g chains. It 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 pre-initiation 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. Its activity is regulated by phosphorylation at serine 48 and serine 51 (without methionine), which stabilizes the eIF-2/GDP/eIF-2B complex and prevents the GDP/GTP exchange reaction, thus impairing the recycling of eIF-2a between successive rounds of initiation and leading to global inhibition of translation. elF-2a is reported to be a substrate for eukaryotic translation initiation factor 2-alpha kinase 1 to 4 (EIF2AK1, 2, 3, and 4). It is also a key component of the integrated stress response (ISR) and is required for adaptation to various stress conditions. Phosphorylation on serine 51 by the EIF2AK4 is shown to occur in response to amino acid starvation and UV irradiation. Its phosphorylation by EIF2AK1-4 converts it into a global protein synthesis inhibitor, leading to reduction in cap-dependent translation, while concomitantly initiating the preferential translation of ISR-specific mRNAs, such as the transcriptional activator ATF4. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Lee, YY., et al. (2009). J. Biol. Chem. 284(11); 6661-6673; Mikami, S., et al. (2006). Protein Expr, Purif. 46(2); 348-357). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |