Product nameAnti-GRP78 BiP antibody
See all GRP78 BiP primary antibodies
DescriptionRabbit polyclonal to GRP78 BiP
Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
Species reactivityReacts with: Mouse, Chicken, Human
Predicted to work with: Rat, Hamster, Xenopus laevis
Synthetic peptide within Human GRP78 BiP aa 1-100. The exact sequence is proprietary.
Database link: P11021
- HeLa cells, breast carcinoma.
This product is FOR RESEARCH USE ONLY. For commercial use, please contact firstname.lastname@example.org.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.60
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab32618 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Detects a band of approximately 75 kDa (predicted molecular weight: 78 kDa).|
|IHC-P||1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
FunctionProbably plays a role in facilitating the assembly of multimeric protein complexes inside the endoplasmic reticulum. Involved in the correct folding of proteins and degradation of misfolded proteins via its interaction with DNAJC10, probably to facilitate the release of DNAJC10 from its substrate.
Involvement in diseaseAutoantigen in rheumatoid arthritis.
Sequence similaritiesBelongs to the heat shock protein 70 family.
Cellular localizationEndoplasmic reticulum lumen. Melanosome. Cytoplasm. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
- Information by UniProt
- 78 kDa glucose regulated protein antibody
- 78 kDa glucose-regulated protein antibody
- AL022860 antibody
All lanes : Anti-GRP78 BiP antibody (ab32618) at 1 µg/ml
Lane 1 : Liver (Mouse) Tissue Lysate
Lane 2 : CHO-K1 cell lysate Whole Cell Lysate
Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 78 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 30 seconds
The band observed at 75 kDa could potentially be a cleaved form of GRP78 BiP due to the presence of a 18 amino acid signal peptide.
ICC/IF image of ab32618 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32618, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
This image shows human breast carcinoma stained with ab32618 diluted 1/100.
Ab32618 staining Human normal liver parenchyma. Staining is localised to endoplasmic reticulum compartment.
Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be requi
This product has been referenced in:
- Bandla S et al. ATPase activity of human binding immunoglobulin protein (BiP) variants is enhanced by signal sequence and physiological concentrations of Mn2. FEBS Open Bio 9:1355-1369 (2019). Read more (PubMed: 31033254) »
- Guan G et al. Mechanism of interactions between endoplasmic reticulum stress and autophagy in hypoxia/reoxygenation-induced injury of H9c2 cardiomyocytes. Mol Med Rep 20:350-358 (2019). Read more (PubMed: 31115545) »