Anti-GRP94 Monoclonal Antibody

Product Name

Anti-GRP94 Monoclonal Antibody

View all gp96/HSP90B1/GRP94 Antibodies

SKU/Catalog Number

M02393-2

Size

200ug

Form

liquid

Description

Boster Bio Anti-GRP94 Monoclonal Antibody catalog # M02393-2. Tested in IP, IF, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20°C for one year. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-GRP94 Monoclonal Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # M02393-2)

Host

Rat

Contents

PBS pH7.2, 50% glycerol, 0.09% sodium azide

Clonality

Monoclonal

Clone Number

9G10

Isotype

IgG2a

Immunogen

Purified Grp94 isolated from chicken oviducts

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

Detects ~98kDa. Does not detect human HSP90, Grp74, or GrpE from E.coli.

Reactive Species

M02393-2 is reactive to HSP90B1 in Bovine, Chicken, Dog, Guinea pig, Hamster, Human, Monkey, Mouse, Pig, Rabbit, Rat, Sheep, Xenopus, Horse

Applications

M02393-2 is guaranteed for Flow Cytometry, IP, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

Calculated molecular weight

92.469kDa

Background of gp96/HSP90B1/GRP94

Grp94 (glucose regulated protein 94, gp96) is a constitutively expressed endoplasmic reticulum (ER) lumenal protein that is up-regulated in response to cellular stress such as heat shock, oxidative stress or glucose depletion. Grp94 is thought to play a role in protein translocation to the ER, in their subsequent folding and assembly, and in regulating protein secretion (1). Grp94 also plays a role in antigen presentation by accessing the endogenous pathway and eliciting specific CTL responses to chaperone bound peptides via MHC class I pathway (2). Grp94 is a member of the HSP90 family of stress proteins and shares sequence homology with its cytosolic equivalent, HSP90 (3). Both HSP90 and Grp94 are calcium binding proteins (4). Despite sharing 50% sequence homology over its N domains and complete conservation in its ligand binding domains with HSP90, Grp94 and HSP90 differ in their interactions with regulatory ligands as Grp94 has weak ATP binding and hydrolysis activity (5). Grp94 exists as a homodimer and the two subunits interact at two distinct intermolecular sites, C terminal dimerization domains and the N-terminal interacts with the middle domain of opposing subunits (6). Grp94 contains a carboxy terminal KDEL (Lys-Asp-Glu-Leu) sequence which is believed to aid in its retention in the ER (7).

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

WB (1:2000), ICC/IF (1:100); optimal dilutions for assays should be determined by the user.

Validation Images & Assay Conditions

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Figure 3. Western blot analysis of HSP90B1 using anti-HSP90B1 antibody (M02393-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-HSP90B1 antigen affinity purified polyclonal antibody (Catalog # M02393-2) at 0.5 ug/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-Rat IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # SA1025) with Tanon 5200 system. A specific band was detected for HSP90B1.

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Immunocytochemistry/Immunofluorescence analysis using Rat Anti-GRP94 Monoclonal Antibody, Clone 9G10 (M02393-2) . Tissue: Heat Shocked HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rat Anti-GRP94 Monoclonal Antibody (M02393-2) at 1:100 for 12 hours at 4°C. Secondary Antibody: R-PE Goat Anti-Rat (yellow) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Endoplasmic reticulum lumen. Melanosome. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-GRP94 Antibody. (C) Composite. Heat Shocked at 42°C for 1h.

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Immunocytochemistry/Immunofluorescence analysis using Rat Anti-GRP94 Monoclonal Antibody, Clone 9G10 (M02393-2) . Tissue: Heat Shocked HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rat Anti-GRP94 Monoclonal Antibody (M02393-2) at 1:100 for 12 hours at 4°C. Secondary Antibody: APC Goat Anti-Rat (red) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Endoplasmic reticulum lumen. Melanosome. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-GRP94 Antibody. (C) Composite. Heat Shocked at 42°C for 1h.

Gene/Protein Information For HSP90B1 (Source: Uniprot.org, NCBI)

Alternative Names

94 kDa glucose-regulated protein; ECGP; Endoplasmin; endothelial cell (HBMEC) glycoprotein; glucose regulated protein, 94 kDa; gp96; GP96ECGP; Grp94; GRP-94; GRP94gp96 homolog; Heat shock protein 90 kDa beta member 1; heat shock protein 90kDa beta (Grp94), member 1; HSP90B1; TRA1endoplasmin; tumor rejection antigen (gp96) 1; Tumor rejection antigen 1; Tumor rejection antigen-1 (gp96) HSP90B1 ECGP, GP96, GRP94, HEL-S-125m, HEL35, TRA1 heat shock protein 90 beta family member 1 endoplasmin|94 kDa glucose-regulated protein|endothelial cell (HBMEC) glycoprotein|epididymis luminal protein 35|epididymis secretory sperm binding protein Li 125m|heat shock protein 90 kDa beta member 1|heat shock protein 90kDa beta (Grp94), member 1|heat shock protein 90kDa beta family member 1|stress-inducible tumor rejection antigen gp96|tumor rejection antigen (gp96) 1|tumor rejection antigen 1

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