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Products:Tags & Cell Markers >> Subcellular Markers >> Organelles >> ER
Overview
Product name
Anti-GRP78 BiP antibody
See all GRP78 BiP products (13) ...
Description
Rabbit polyclonal to GRP78 BiP
Tested applications
WB, IHC-P, ICC/IFmore details
Cross reactivity
Reacts with
Mouse, Human
Predicted to work with
Rat, Chicken, Hamster, Xenopus laevis
Immunogen
Synthetic peptide: KEDVGTV VGIDLGTTYS CVG, corresponding to amino acids 24-43 of Human GRP78 BiP
KEDVGTV VGIDLGTTYS CVG
Positive control
HeLa cells, breast carcinoma.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer
Preservative: 0.1% Sodium Azide
Constituents: 1% BSA, 10mM PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Protein A purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Tags & Cell Markers >> Subcellular Markers >> Organelles >> ER
Applications
Show applications key
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 dilution. Detects a band of approximately 75 kDa (predicted molecular weight: 78 kDa).
IHC-P: 1/100Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF: Use a concentration of 1 µg/ml
Target
Function
Probably plays a role in facilitating the assembly of multimeric protein complexes inside the ER.
Involvement in disease
Note=Autoantigen in rheumatoid arthritis.
Sequence similarities
Belongs to the heat shock protein 70 family.
Cellular localization
Endoplasmic reticulum lumen. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
Target information above from: UniProt accessionP11021
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- 78 kDa glucose regulated protein antibody
- 78 kDa glucose-regulated protein antibody
- BIP antibody
see all
Database links
- Entrez Gene: 3309 Human
- Entrez Gene: 14828 Mouse
- Entrez Gene: 25617 Rat
- Entrez Gene: 397850 Xenopus laevis
- Omim: 138120 Human
- SwissProt: P11021 Human
- SwissProt: P20029 Mouse
- SwissProt: P06761 Rat
see all
Anti-GRP78 BiP antibody images:
Microtomy (Formalin-fixed paraffin-embedded sections) - GRP78 BiP antibody (ab32618)
This image shows human breast carcinoma stained with ab32618 diluted 1/100.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - GRP78 BiP antibody (ab32618)
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 required.
Immunocytochemistry/ Immunofluorescence-GRP78 BiP antibody(ab32618)
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.
Western blot - GRP78 BiP antibody (ab32618)
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.
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (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.
References for Anti-GRP78 BiP antibody (ab32618)
This product has been referenced in:
- Cunnea Pet al. Expression profiles of endoplasmic reticulum stress-related molecules in demyelinating lesions and multiple sclerosis. Mult Scler 17:808-18 (2011). IHC-P; Human.Read more (PubMed: 21382862) »
- Huang Yet al. PML-RARa enhances constitutive autophagic activity through inhibiting the Akt/mTOR pathway. Autophagy 7:1132-44 (2011). WB; Human.Read more (PubMed: 21673516) »
See all 6 publications for this product
Publishing research using ab32618? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"