Anti-ERp57 antibody (ab10287)
- Datasheet
- References (20)
- Protocols
Overview
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Product nameAnti-ERp57 antibody
See all ERp57 primary antibodies -
DescriptionRabbit polyclonal to ERp57
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Host speciesRabbit
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Tested applicationsSuitable for: Flow Cyt, IHC-P, ICC/IF, WB, ICC, IPmore details
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Species reactivityReacts with: Mouse, Human
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Immunogen
His-tagged E. coli expressed recombinant ERp57.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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PurityWhole antiserum
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab10287 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Flow Cyt | Use at an assay dependent concentration. PubMed: 23762310 ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody. |
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| IHC-P | Use at an assay dependent concentration. PubMed: 20731657 | |
| ICC/IF | Use at an assay dependent concentration. | |
| WB | 1/1000 - 1/3000. Detects a band of approximately 57 kDa (predicted molecular weight: 61 kDa). | |
| ICC | 1/100. | |
| IP | Use at an assay dependent concentration. |
Target
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Tissue specificityDetected in the flagellum and head region of spermatozoa (at protein level).
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Sequence similaritiesBelongs to the protein disulfide isomerase family.
Contains 2 thioredoxin domains. -
Cellular localizationEndoplasmic reticulum lumen. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
- Information by UniProt
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Database links
- Entrez Gene: 2923 Human
- Entrez Gene: 14827 Mouse
- Omim: 602046 Human
- SwissProt: P30101 Human
- SwissProt: P27773 Mouse
- Unigene: 591095 Human
- Unigene: 263177 Mouse
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Alternative names
- 58 kDa glucose regulated protein antibody
- 58 kDa glucose-regulated protein antibody
- 58 kDa microsomal protein antibody
see all
Images
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Western blot - Anti-ERp57 antibody (ab10287)All lanes : Anti-ERp57 antibody (ab10287) at 1/1000 dilution
Lane 1 : Mouse embryonic cells
Lane 2 : Calreticulin deficient mouse embryomic cells
Predicted band size: 61 kDa
Observed band size: 57 kDa why is the actual band size different from the predicted?Western blot using ab10287 at 1/1000.
Lane 1 – Mouse Embryonic Fibroblasts.
Lane 2 – Calreticulin deficient Mouse Embryonic Fibroblasts. -
Immunocytochemistry/ Immunofluorescence - Anti-ERp57 antibody (ab10287)Double staining of mouse embryonic fibroblasts (top) and calreticulin deficient fibroblasts (bottom) with rabbit-anti-ERp57 antibodies (1:100; upper and lower left panel) and FITC-ConA (upper and lower right panel). -
Immunocytochemistry/ Immunofluorescence - Anti-ERp57 antibody (ab10287)This image is courtesy of an anonymous Abreviewab10287 at 1/100 staining human HeLa cells by ICC/IF. The cells were premeabilized with digitonin before being blocked and then stained with the antibody for 1 hour. An Alexa-Fluor ® conjugated goat anti rabbit antibody was used as the secondary.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERp57 antibody (ab10287)ab10287 (1/2000) staining ERp57 in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining.
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 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required. -
Immunoprecipitation - Anti-ERp57 antibody (ab10287)ERp57 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to ERp57 (ab10287) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab10287.
Secondary: Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 61kDa: ERp57
Protocols
Datasheets and documents
References
This product has been referenced in:
- Mazumder A et al. Hydroxycoumarin OT-55 kills CML cells alone or in synergy with imatinib or Synribo: Involvement of ER stress and DAMP release. Cancer Lett 438:197-218 (2018). Read more (PubMed: 30205168) »
- Stopa JD et al. Kinetic-based trapping by intervening sequence variants of the active sites of protein-disulfide isomerase identifies platelet protein substrates. J Biol Chem 292:9063-9074 (2017). Read more (PubMed: 28364042) »
