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Human GRP78 BiP ELISA Kit is a single-wash 90-min Simplestep used to quantify Human GRP78 BiP with a sensitivity of 0.051 ng/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair


Key facts

Detection method
Colorimetric
Sample types
Cell Lysate, Tissue Extracts
Assay type
Sandwich (quantitative)
Reactive species
Human
Range
0.219 - 14 ng/mL
Assay time
1h 30m
Sensitivity
= 0.051 ng/mL

Reactivity data

Application
sELISA
Reactivity
Reacts
Dilution info
-
Notes

-

Target data

Function

Endoplasmic reticulum chaperone that plays a key role in protein folding and quality control in the endoplasmic reticulum lumen (PubMed:2294010, PubMed:23769672, PubMed:23990668, PubMed:28332555). Involved in the correct folding of proteins and degradation of misfolded proteins via its interaction with DNAJC10/ERdj5, probably to facilitate the release of DNAJC10/ERdj5 from its substrate (By similarity). Acts as a key repressor of the EIF2AK3/PERK and ERN1/IRE1-mediated unfolded protein response (UPR) (PubMed:1550958, PubMed:11907036, PubMed:19538957). In the unstressed endoplasmic reticulum, recruited by DNAJB9/ERdj4 to the luminal region of ERN1/IRE1, leading to disrupt the dimerization of ERN1/IRE1, thereby inactivating ERN1/IRE1 (By similarity). Also binds and inactivates EIF2AK3/PERK in unstressed cells (PubMed:11907036). Accumulation of misfolded protein in the endoplasmic reticulum causes release of HSPA5/BiP from ERN1/IRE1 and EIF2AK3/PERK, allowing their homodimerization and subsequent activation (PubMed:11907036). Plays an auxiliary role in post-translational transport of small presecretory proteins across endoplasmic reticulum (ER). May function as an allosteric modulator for SEC61 channel-forming translocon complex, likely cooperating with SEC62 to enable the productive insertion of these precursors into SEC61 channel. Appears to specifically regulate translocation of precursors having inhibitory residues in their mature region that weaken channel gating. May also play a role in apoptosis and cell proliferation (PubMed:26045166). (Microbial infection) Plays an important role in viral binding to the host cell membrane and entry for several flaviruses such as Dengue virus, Zika virus and Japanese encephalitis virus (PubMed:15098107, PubMed:28053106, PubMed:33432092). Acts as a component of the cellular receptor for Dengue virus serotype 2/DENV-2 on human liver cells (PubMed:15098107). (Microbial infection) Acts as a receptor for CotH proteins expressed by fungi of the order mucorales, the causative agent of mucormycosis, which plays an important role in epithelial cell invasion by the fungi (PubMed:20484814, PubMed:24355926, PubMed:32487760). Acts as a receptor for R.delemar CotH3 in nasal epithelial cells, which may be an early step in rhinoorbital/cerebral mucormycosis (RCM) disease progression (PubMed:32487760).

Alternative names

What's included?

96 Tests
Components
10X Human GRP78 BiP Capture Antibody
1 x 600 µL
10X Human GRP78 BiP Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
50X Cell Extraction Enhancer Solution (ab193971)
1 x 1 mL
5X Cell Extraction Buffer PTR (ab193970)
1 x 10 mL
Antibody Diluent CPR2
1 x 6 mL
Human GRP78 BiP Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seal
1 x 1 Unit
Pre-Coated 96-Well Microplate
1 x 96 Wells
Sample Diluent NS (ab193972)
1 x 12 mL
Stop Solution
1 x 12 mL
TMB Development Solution
1 x 12 mL

Recommended products

Human GRP78 BiP ELISA Kit is a single-wash 90-min Simplestep used to quantify Human GRP78 BiP with a sensitivity of 0.051 ng/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Design your own immunoassay: we also offer the conjugation-ready antibody pair

Key facts

Detection method
Colorimetric
Sample types
Cell Lysate, Tissue Extracts
Assay type
Sandwich (quantitative)
Reactive species
Human
Range
0.219 - 14 ng/mL
Assay time
1h 30m
Assay Platform
Pre-coated microplate (12 x 8 well strips)
Sensitivity
= 0.051 ng/mL

Precision

Intra assay

Sample
Extract
n
8
C.V.
7.5

Inter assay

Sample
Extract
n
3
C.V.
9.1

Recovery

Sample specific recovery

Sample type
Cell Lysate
Average %
= 106
Range
87 - 123 %
Sample type
Tissue Extracts
Average %
= 82
Range
78 - 87 %

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C

Notes

Human GRP78 BiP SimpleStep ELISA® kit is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of Human GRP78 BiP protein in human cell extract and tissue extract. Quantitate Human GRP78 BiP with 0.051 ng/ml sensitivity.

SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:

-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips

A 384-well SimpleStep ELISA® microplate (Pre-coated 384 well Microplate SimpleStep ELISA® ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

GRP78 also known as BiP or HSPA5 is a protein that plays an important role in protein folding and assembly within the endoplasmic reticulum (ER). It weighs approximately 78 kDa hence the name GRP78. This chaperone protein binds to hydrophobic regions of nascent polypeptides to prevent aggregation and misfolding. GRP78 is expressed in high levels in the ER of cells where it monitors cellular stress and aids in maintaining ER homeostasis.

Biological function summary

GRP78/BiP is important in the unfolded protein response (UPR) a cellular stress response related to the ER. It is part of a complex that manages protein load inside the ER by regulating the fold of nascent proteins and interacting with other ER stress sensors like IRE1 PERK and ATF6. This function is essential for maintaining proper protein conformation in the ER especially during physiological stress ensuring that only correctly folded proteins proceed to the Golgi apparatus.

Pathways

GRP78/BiP significantly affects the UPR and apoptosis pathways. By controlling protein folding and quality in the ER GRP78 helps prevent cell death under stress conditions. It works closely with other proteins like ATF6 which activates stress response genes. In normal conditions GRP78 keeps stress transducers inactivated but during stress it dissociates allowing UPR signaling to occur.

Associated diseases and disorders

GRP78/BiP has been implicated in cancer and neurodegenerative diseases. Overexpression of GRP78 is associated with tumor proliferation and poor prognosis in various cancers as cancer cells heavily rely on its protein-folding capacity to survive. Additionally in neurodegenerative diseases misfolded proteins can accumulate leading GRP78 to attempt counteracting these toxic aggregations highlighting its connection with proteins like tau and amyloid-beta in diseases such as Alzheimer's.

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