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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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- Product number
- 701268 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-EIF2S1 (Ser51) Recombinant Rabbit Monoclonal Antibody (10H21L20)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with mouse, rat, non-human primate, rabbit and equine based on sequence homology.
- Antibody clone number
- 10H21L20
- Concentration
- 0.5 mg/mL
Submitted references Integrated Stress Response Activity Marks Stem Cells in Normal Hematopoiesis and Leukemia.
van Galen P, Mbong N, Kreso A, Schoof EM, Wagenblast E, Ng SWK, Krivdova G, Jin L, Nakauchi H, Dick JE
Cell reports 2018 Oct 30;25(5):1109-1117.e5
Cell reports 2018 Oct 30;25(5):1109-1117.e5
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Phospho-eIF2a pSer51 in whole cell extracts from K562 cells treated with IFN alpha (100 ng/mL, 20 min) using a Phospho-eIF2a pSer51 recombinant rabbit monoclonal antibody (Product # 701268) at a dilution of 2 µg/mL. To confirm specificity, competition was performed by preincubation with the phosphopeptide to inhibit antibody binding (lane 2). Samples were detected using chemiluminescence (ECL). Results show a band at ~36 kDa.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Phospho-eIF2a pSer51 in HeLa cells using a Phospho-eIF2a pSer51 recombinant rabbit monoclonal antibody (Product # 701268) followed by detection using an Alexa Fluor 488-conjugated goat anti-rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor 594 phalloidin (red) (image C). Image D is a composite image showing nuclear localization of EIF2a and Image E is a composite image of cells showing inhibition of antibody binding after competition with the phosphopeptide.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of EIF2A (PS51) showing staining in the cytoplasm of paraffin-embedded human breast carcinoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a EIF2A Recombinant Rabbit Monoclonal Antibody (Product # 701268) diluted in 3% BSA-PBS at a dilution of 1:20 for 1 hour at 37ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
