Anti-GRP94/HSP90B1 Antibody

Product Name

Anti-GRP94/HSP90B1 Antibody

View all gp96/HSP90B1/GRP94 Antibodies

SKU/Catalog Number

PA1340

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-GRP94/HSP90B1 Antibody catalog # PA1340. Tested in IHC, ICC, WB applications. This antibody reacts with Human, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-GRP94/HSP90B1 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1340)

Host

Rabbit

Contents

Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence in the middle region of human GRP94, identical to the related mouse and rat sequences.

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

Cross-reactivity

No cross-reactivity with other proteins

Reactive Species

PA1340 is reactive to HSP90B1 in Human, Rat

Applications

PA1340 is guaranteed for IHC, IHC-F, ICC, WB Boster Guarantee

Observed Molecular Weight

100 kDa

Calculated molecular weight

92.469kDa

Background of gp96/HSP90B1/GRP94

Heat shock protein 90kDa beta member 1  (HSP90B1), known also as endoplasmin, GRP94, is a chaperone protein that in humans is encoded by the HSP90B1 gene. It is mapped to chromosome 12q23.3. This gene encodes a member of a family of adenosine triphosphate (ATP)-metabolizing molecular chaperones with roles in stabilizing and folding other proteins. The encoded protein is localized to melanosomes and the endoplasmic reticulum. Expression of this protein is associated with a variety of pathogenic states, including tumor formation.

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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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

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.

Western blot, 0.1-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Rat, By Heat
Immunohistochemistry (Frozen Section), 0.5-1μg/ml, Rat
Immunocytochemistry , 0.5-1μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of GRP94/HSP90B1 using anti-GRP94/HSP90B1 antibody (PA1340).
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 30 ug of sample under reducing conditions.
Lane 1: human A549 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human Jurkat whole cell lysates,
Lane 4: human K562 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-GRP94/HSP90B1 antigen affinity purified polyclonal antibody (Catalog # PA1340) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GRP94/HSP90B1 at approximately 100 kDa. The expected band size for GRP94/HSP90B1 is at 92 kDa.

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Figure 2. ICC analysis of GRP94/HSP90B1 using anti-GRP94/HSP90B1 antibody (PA1340).
GRP94/HSP90B1 was detected in an immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 1 μg/ml rabbit anti-GRP94/HSP90B1 Antibody (PA1340) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. IHC analysis of GRP94/HSP90B1 using anti-GRP94/HSP90B1 antibody (PA1340).
GRP94/HSP90B1 was detected in a paraffin-embedded section of Human Lung Cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-GRP94/HSP90B1 Antibody (PA1340) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 4. IHC analysis of GRP94/HSP90B1 using anti-GRP94/HSP90B1 antibody (PA1340).
GRP94/HSP90B1 was detected in a paraffin-embedded section of Rat Brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-GRP94/HSP90B1 Antibody (PA1340) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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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