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GRP78/HSPA5 Antibody (C9-9-R)

Novus Biologicals, part of Bio-Techne | Catalog # NBP3-32408

Recombinant Monoclonal Antibody
Novus Biologicals, part of Bio-Techne
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NBP3-32408

Key Product Details

Species Reactivity

Human, Mouse, Rat, Zebrafish

Applications

Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Mouse IgG1 Clone # C9-9-R

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Product Specifications

Immunogen

Recombinant protein within human GRP78 aa 100-400. (Uniprot: P11021)

Localization

Cytoplasm, endoplasmic reticulum lumen

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

Theoretical MW

72 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for GRP78/HSPA5 Antibody (C9-9-R)

GRP78/HSPA5 Antibody (C9-9-R)

Western Blot: GRP78/HSPA5 Antibody (C9-9-R) [NBP3-32408]

Western Blot: GRP78/HSPA5 Antibody (C9-9-R) [NBP3-32408] - Western blot analysis of GRP78/HSPA5 on different lysates with Mouse anti-GRP78/HSPA5 antibody (NBP3-32408) at 1/1,000 dilution.

Lane 1: MCF7 cell lysate (15 ug/Lane)
Lane 2: HepG2 cell lysate (15 ug/Lane)
Lane 3: Mouse brain tissue lysate (30 ug/Lane)
Lane 4: U-87 MG cell lysate (30 ug/Lane)
Lane 5: RAW264.7 cell lysate (30 ug/Lane)
Lane 6: RAW264.7 treated with 300nM Thapsigargin for 18 hours cell lysate (30 ug/Lane)
Lane 7: Mouse liver tissue lysate (30 ug/Lane)
Lane 8: Rat liver tissue lysate (30 ug/Lane)
Lane 9: Rat pancreas tissue lysate (30 ug/Lane)

Predicted band size: 72 kDa
Observed band size: 72 kDa

Exposure time: Lane 1-3: 11 seconds; Lane 4-9: 4 seconds; ECL;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (NBP3-32408) at 1/1,000 dilution was used in 5% NFDM/TBST at 4 overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
GRP78/HSPA5 Antibody (C9-9-R)

Immunohistochemistry: GRP78/HSPA5 Antibody (C9-9-R) [NBP3-32408]

Immunohistochemistry: GRP78/HSPA5 Antibody (C9-9-R) [NBP3-32408] - Immunohistochemical analysis of paraffin-embedded rat brain tissue with Mouse anti-GRP78/HSPA5 antibody (NBP3-32408) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (NBP3-32408) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
GRP78/HSPA5 Antibody (C9-9-R)

Immunocytochemistry/ Immunofluorescence: GRP78/HSPA5 Antibody (C9-9-R) [NBP3-32408]

Immunocytochemistry/ Immunofluorescence: GRP78/HSPA5 Antibody (C9-9-R) [NBP3-32408] - Immunocytochemistry analysis of Hela cells labeling GRP78/HSPA5 with Mouse anti-GRP78/HSPA5 antibody (NBP3-32408) at 1/200 dilution.

Cells were fixed in 100% methanol for 10 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-GRP78/HSPA5 antibody (NBP3-32408) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

beta Tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594) were used as the secondary antibody at 1/1,000 dilution.

Applications for GRP78/HSPA5 Antibody (C9-9-R)

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:200

Immunohistochemistry-Paraffin

1:200

Western Blot

1:1000
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS (pH7.4), 0.1% BSA and 40% Glycerol

Preservative

0.05% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: GRP78/HSPA5

Within the ER, misfolded proteins are detected by Bip (GPR78). In the presence of misfolded proteins, GPR78 disscociates from PERK, allowing it to homodimerize and autophosphoyate. In its dimerized phosphorylated form, PERK phosphorylates eIF2. The phosphorylation of eIF2 blocks the majority of translation to prevent the continued accumulation of protein in the ER. GRP78 also binds to IRE1 and ATF6 under normal ER conditions, but dissociates upon accumulation of misfolded proteins. Unbound ATF6 is translocated to the gogli where it is converted into a cleaved, active form. The cleaved form of ATF6 is then able to up regulate transcription of UPR genes including XBP1. Activated IRE1 acts as an endoribonuclease to an intron from XBP1 transcripts. The XBP1 splice variant codes for an active transcription factor which activates transcription of P58IPK. P58IPK is a HSP40 protein which binds to and inhibits PERK. Thus, if the accumulated misfolded proteins have been removed, P58IPK acts to shut down the UPR by removing the block to translation.

Long Name

75 KD Glucose Regulated Protein

Alternate Names

BIP, HSP70-5, HSPA5

Gene Symbol

HSPA5

Additional GRP78/HSPA5 Products

Product Documents for GRP78/HSPA5 Antibody (C9-9-R)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for GRP78/HSPA5 Antibody (C9-9-R)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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