Recombinant Anti-GRP94 antibody [EPR22847-50] (ab238126)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22847-50] to GRP94
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-GRP94 antibody [EPR22847-50]
See all GRP94 primary antibodies -
Description
Rabbit monoclonal [EPR22847-50] to GRP94 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human kidney and stomach tissue lysates; mouse brain and placenta tissue lysates; rat brain, placenta and stomach tissue lysates; HeLa, MCF7, HEK293T, MEF, PC-12, C6 and NIH/3T3 whole cell lysates. IHC-P: Human kidney tissue; mouse and rat colon tissue. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22847-50 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab238126 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 |
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WB |
1/1000. Detects a band of approximately 94 kDa (predicted molecular weight: 94 kDa).
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IHC-P |
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/1000.
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IP |
1/30.
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Flow Cyt (Intra) |
1/600.
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Notes |
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WB
1/1000. Detects a band of approximately 94 kDa (predicted molecular weight: 94 kDa). |
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/1000. |
IP
1/30. |
Flow Cyt (Intra)
1/600. |
Target
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Function
Molecular chaperone that functions in the processing and transport of secreted proteins. Functions in endoplasmic reticulum associated degradation (ERAD). Has ATPase activity. -
Sequence similarities
Belongs to the heat shock protein 90 family. -
Cellular localization
Endoplasmic 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: 7184 Human
- Entrez Gene: 22027 Mouse
- Entrez Gene: 362862 Rat
- Omim: 191175 Human
- SwissProt: P14625 Human
- SwissProt: P08113 Mouse
- SwissProt: Q66HD0 Rat
- Unigene: 192374 Human
see all -
Alternative names
- 94 kDa glucose regulated protein antibody
- 94 kDa glucose-regulated protein antibody
- ECGP antibody
see all
Images
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All lanes : Anti-GRP94 antibody [EPR22847-50] (ab238126) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : HSP90B1 knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 94 kDa
Observed band size: 94 kDaLanes 1-3: Merged signal (red and green). Green - ab238126 observed at 94 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab238126 Anti-GRP94 antibody [EPR22847-50] was shown to specifically react with GRP94 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266313 (knockout cell lysate ab257254) was used. Wild-type and GRP94 knockout samples were subjected to SDS-PAGE. ab238126 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] (ab238126)
Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling GRP94 with ab238126 at 1/5000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on rat colon (PMID: 23572575) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
The section was incubated with ab238126 for 15 mins at RT.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] (ab238126)
Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling GRP94 with ab238126 at 1/5000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on mouse colon (PMID: 23572575) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
The section was incubated with ab238126 for 15 mins at RT.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] (ab238126)
Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling GRP94 with ab238126 at 1/5000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human kidney (PMID: 20520781) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
The section was incubated with ab238126 for 15 mins at RT.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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GRP94 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab238126 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab238126 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.
Lane 1: HeLa whole cell lysate 10 μg (Input).
Lane 2: ab238126 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab238126 in HeLa whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds. -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling GRP94 with ab238126 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue).
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-GRP94 antibody [EPR22847-50] (ab238126) at 1/1000 dilution
Lane 1 : Mouse brain tissue lysate at 20 µg
Lane 2 : Mouse placenta tissue lysate at 20 µg
Lane 3 : Mouse stomach tissue lysate at 20 µg
Lane 4 : Rat brain tissue lysate at 20 µg
Lane 5 : Rat placenta tissue lysate at 20 µg
Lane 6 : Rat stomach tissue lysate at 20 µg
Lane 7 : C6 (rat glial tumor glial cell line) whole cell lysate at 20 µg
Lane 8 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 94 kDa
Observed band size: 94 kDaBlocking and dilution buffer: 5% NFDM/TBST.
Exposure times.
Lane 1: 26 seconds; Lanes 2-5: 10 seconds; Lane 6: 92 seconds; Lanes 7-8: 6 seconds.
The molecular weight observed is consistent with what has been described in the literature (PMID: 26858453, 29949743).
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All lanes : Anti-GRP94 antibody [EPR22847-50] (ab238126) at 1/1000 dilution
Lane 1 : Human brain tissue lysate at 20 µg
Lane 2 : Human kidney tissue lysate at 20 µg
Lane 3 : Human stomach tissue lysate at 20 µg
Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 5 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 6 : MEF (Mouse embryonic fibroblast (immortalized) cell line) whole cell lysate at 20 µg
Lane 7 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 94 kDa
Observed band size: 94 kDaBlocking and dilution buffer: 5% NFDM/TBST.
Exposure times.
Lanes 1 & 2: 10 seconds; Lane 3: 37 seconds; Lanes 4-6: 10 seconds; Lane 7: 26 seconds.
The molecular weight observed is consistent with what has been described in the literature (PMID: 26858453, 29949743).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling GRP94 with ab238126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing endoplasmic reticulum staining in NIH/3T3 cell line. The nuclear counterstain is DAPI (blue). The endoplasmic reticulum is stained with Anti-KDEL antibody [10C3] (ab12223), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) secondary antibody (red).
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling GRP94 with ab238126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing endoplasmic reticulum staining in HeLa cell line. The nuclear counter stain is DAPI (blue). The endoplasmic reticulum is stained with Anti-KDEL antibody [10C3] (ab12223), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) secondary antibody (red).
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (4)
ab238126 has been referenced in 4 publications.
- Zhang Y et al. Phosphatase Shp2 regulates biogenesis of small extracellular vesicles by dephosphorylating Syntenin. J Extracell Vesicles 10:e12078 (2021). PubMed: 33732417
- Persaud AK et al. Facilitative lysosomal transport of bile acids alleviates ER stress in mouse hematopoietic precursors. Nat Commun 12:1248 (2021). PubMed: 33623001
- Mo C et al. LncRNA nuclear-enriched abundant transcript 1 shuttled by prostate cancer cells-secreted exosomes initiates osteoblastic phenotypes in the bone metastatic microenvironment via miR-205-5p/runt-related transcription factor 2/splicing factor proline- and glutamine-rich/polypyrimidine tract-binding protein 2 axis. Clin Transl Med 11:e493 (2021). PubMed: 34459124
- Jiang K et al. Exosome-derived ENO1 regulates integrin a6ß4 expression and promotes hepatocellular carcinoma growth and metastasis. Cell Death Dis 11:972 (2020). PubMed: 33184263