GRP94 Polyclonal antibody

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  WB analysis using 14700-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 14700-1-AP (GRP94 antibody) at dilution of 1:8000 incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 14700-1-AP

  HEK-293 cells and HEK-293-derived exosomes (HEK-293-exo) were subjected to SDS PAGE followed by western blot with 14700-1-AP (GRP94 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IP experiment of mouse liver using 14700-1-AP

  IP result of anti-GRP94 (IP:14700-1-AP, 3ug; Detection:14700-1-AP 1:1000) with mouse liver tissue lysate 4000ug.

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  IP experiment of mouse liver using 14700-1-AP

  IP result of anti-GRP94 (IP:14700-1-AP, 4ug; Detection:14700-1-AP 1:2000) with mouse liver tissue lysate 1560 ug.

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  IHC staining of human breast cancer using 14700-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 14700-1-AP (GRP94 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 14700-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 14700-1-AP (GRP94 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human cervical cancer using 14700-1-AP

  Immunohistochemical analysis of paraffin-embedded human cervical cancer slide using 14700-1-AP (HSP90B1 Antibody) at dilution of 1:50.

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  IHC staining of human cervical cancer using 14700-1-AP

  Immunohistochemical analysis of paraffin-embedded human cervical cancer slide using 14700-1-AP (HSP90B1 Antibody) at dilution of 1:50.

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  IF Staining of HeLa using 14700-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using GRP94 antibody (14700-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

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  IF Staining of U2OS using 14700-1-AP

  Immunofluorescent analysis of (4% PFA) fixed U2OS cells using GRP94 antibody (14700-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-phalloidin (red).

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  IF Staining of HeLa using 14700-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using GRP94 antibody (14700-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-phalloidin (red).

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  IF Staining of HepG2 using 14700-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using GRP94 antibody (14700-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  IF Staining of HepG2 using 14700-1-AP

  Immunofluorescent analysis of HepG2 cells, using HSP90B1 antibody 14700-1-AP at 1:25 dilution and Rhodamine-labeled goat anti-rabbit IgG (red).

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  FC experiment of HeLa using 14700-1-AP

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human GRP94 (14700-1-AP) and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer.

• Featured Product
• KD/KO Validated

GRP94 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Cat No. 14700-1-AP

Publications(52)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA

Synonyms

HSP90B1, 94 kDa glucose-regulated protein, ECGP, Endoplasmin, GP96

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Tested Applications

• Positive WB detected in: A431 cells, HEK-293 cells, A549 cells, HeLa cells, HepG2 cells, MCF-7 cells, U2OS cells
• Positive IP detected in: mouse liver tissue
• Positive IHC detected in: human breast cancer tissue, human cervical cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells, HepG2 cells, U2OS cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:16000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 2 publications below
• WB: See 47 publications below
• IHC: See 6 publications below
• IF: See 3 publications below
• CoIP: See 1 publications below

Product Information

14700-1-AP targets GRP94 in WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen:

  CatNo: Ag6344

  Product name: Recombinant human GRP94 protein

  Source: e coli.-derived, PET28a

  Tag: 6*His

  Domain: 455-803 aa of BC066656

  Sequence: KLLKVIRKKLVRKTLDMIKKIADDKYNDTFWKEFGTNIKLGVIEDHSNRTRLAKLLRFQSSHHPTDITSLDQYVERMKEKQDKIYFMAGSSRKEAESSPFVERLLKKGYEVIYLTEPVDEYCIQALPEFDGKRFQNVAKEGVKFDESEKTKESREAVEKEFEPLLNWMKDKALKDKIEKAVVSQRLTESPCALVASQYGWSGNMERIMKAQAYQTGKDISTNYYASQKKTFEINPRHPLIRDMLRRIKEDEDDKTVLDLAVVLFETATLRSGYLLPDTKAYGDRIERMLRLSLNIDPDAKVEEEPEEEPEETAEDTTEDTEQDEDEEMDVGTDEEEETAKESTAEKDEL

• Full Name: heat shock protein 90kDa beta (Grp94), member 1
• Calculated Molecular Weight: 92 kDa
• Observed Molecular Weight: 100 kDa
• GenBank Accession Number: BC066656
• Gene Symbol: GRP94
• Gene ID (NCBI): 7184
• RRID: AB_2233347
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: P14625
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Glucose-regulated protein 94 (GRP94), also called HSP90B1 and GP96, is an endoplasmic reticulum (ER)-resident member of the heat shock protein 90 (HSP90) family (PMID: 33802964; 24658275). Under ER stress conditions, GRP94 accelerates its function as a molecular chaperone. Client proteins of GRP94 include toll-like receptors (TLRs), glycoprotein (GP) IX subunit, insulin-like growth factors (IGFs), proinsulin, and integrins (PMID: 7913987; 32781621; 22079671). As well as being an ER chaperone, GRP94 can also participate in Calcium regulation and is also a regulator of innate and adaptive immunity (PMID: 33802964; 11584270).

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
human	WB	Cell; A Translocation Pathway for Vesicle-Mediated Unconventional Protein Secretion.; Authors - Min Zhang
human	WB	Sci Adv; Platelet P-selectin initiates cross-presentation and dendritic cell differentiation in blood monocytes.; Authors - Patrick Han
mouse	WB	EMBO J; Rab2A-mediated Golgi-lipid droplet interactions support very-low-density lipoprotein secretion in hepatocytes; Authors - Min Xu
human	WB,IF	Theranostics; Exosomes from mesenchymal stem cells modulate endoplasmic reticulum stress to protect against nucleus pulposus cell death and ameliorate intervertebral disc degeneration in vivo.; Authors - Zhiwei Liao
rat	WB	Redox Biol; Exogenous spermine attenuates rat diabetic cardiomyopathy via suppressing ROS-p53 mediated downregulation of calcium-sensitive receptor.; Authors - Yuehong Wang
human	WB	Haematologica; Comparative analysis of ChAdOx1 nCoV-19 and Ad26.COV2.S SARS-CoV-2 vector vaccines.; Authors - Stephan Michalik