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Human GBA ELISA Kit is a single-wash 90-min Simplestep used to quantify Human GBA with a sensitivity of 10.841 pg/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
- KO validated for guaranteed specificity

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Images

Sandwich ELISA - Human GBA ELISA Kit (AB315298), expandable thumbnail
  • Sandwich ELISA - Human GBA ELISA Kit (AB315298), expandable thumbnail
  • Sandwich ELISA - Human GBA ELISA Kit (AB315298), expandable thumbnail

Key facts

Detection method
Colorimetric
Sample types
Cell Lysate, Tissue Extracts
Assay type
Sandwich (quantitative)
Reactive species
Human
Range
87.5 - 5600 pg/mL
Assay time
1h 30m
Sensitivity
= 10.841 pg/mL

Reactivity data

Application
sELISA
Reactivity
Reacts
Dilution info
-
Notes

-

Target data

Function

Glucosylceramidase that catalyzes, within the lysosomal compartment, the hydrolysis of glucosylceramides/GlcCers (such as beta-D-glucosyl-(1<->1')-N-acylsphing-4-enine) into free ceramides (such as N-acylsphing-4-enine) and glucose (PubMed:15916907, PubMed:24211208, PubMed:32144204, PubMed:9201993). Plays a central role in the degradation of complex lipids and the turnover of cellular membranes (PubMed:27378698). Through the production of ceramides, participates in the PKC-activated salvage pathway of ceramide formation (PubMed:19279011). Catalyzes the glucosylation of cholesterol, through a transglucosylation reaction where glucose is transferred from GlcCer to cholesterol (PubMed:24211208, PubMed:26724485, PubMed:32144204). GlcCer containing mono-unsaturated fatty acids (such as beta-D-glucosyl-N-(9Z-octadecenoyl)-sphing-4-enine) are preferred as glucose donors for cholesterol glucosylation when compared with GlcCer containing same chain length of saturated fatty acids (such as beta-D-glucosyl-N-octadecanoyl-sphing-4-enine) (PubMed:24211208). Under specific conditions, may alternatively catalyze the reverse reaction, transferring glucose from cholesteryl 3-beta-D-glucoside to ceramide (Probable) (PubMed:26724485). Can also hydrolyze cholesteryl 3-beta-D-glucoside producing glucose and cholesterol (PubMed:24211208, PubMed:26724485). Catalyzes the hydrolysis of galactosylceramides/GalCers (such as beta-D-galactosyl-(1<->1')-N-acylsphing-4-enine), as well as the transfer of galactose between GalCers and cholesterol in vitro, but with lower activity than with GlcCers (PubMed:32144204). Contrary to GlcCer and GalCer, xylosylceramide/XylCer (such as beta-D-xyosyl-(1<->1')-N-acylsphing-4-enine) is not a good substrate for hydrolysis, however it is a good xylose donor for transxylosylation activity to form cholesteryl 3-beta-D-xyloside (PubMed:33361282).

Alternative names

What's included?

1 x 96 Tests
Components
10X Human GBA Capture Antibody
1 x 600 µL
10X Human GBA Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
5X Cell Extraction Buffer PTR (ab193970)
1 x 10 mL
Antibody Diluent 4BR
1 x 6 mL
Human GBA Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seals
1 x 1 Unit
Sample Diluent NS (ab193972)
1 x 12 mL
SimpleStep Pre-Coated 96-Well Microplate (ab206978)
1 x 1 Unit
Stop Solution
1 x 12 mL
TMB Development Solution
1 x 12 mL

Recommended products

Human GBA ELISA Kit is a single-wash 90-min Simplestep used to quantify Human GBA with a sensitivity of 10.841 pg/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
- KO validated for guaranteed specificity

Key facts

Detection method
Colorimetric
Sample types
Cell Lysate, Tissue Extracts
Assay type
Sandwich (quantitative)
Reactive species
Human
Range
87.5 - 5600 pg/mL
Assay time
1h 30m
Assay Platform
Pre-coated microplate (12 x 8 well strips)
Sensitivity
= 10.841 pg/mL
General recovery
100%

Precision

Intra assay

Sample
Extract
n
8
C.V.
= 4.4

Inter assay

Sample
Extract
n
3
C.V.
= 8.9

Recovery

Sample specific recovery

Sample type
Cell Lysate
Average %
= 100
Range
94 - 108 %
Sample type
Tissue Extracts
Average %
= 107
Range
88 - 119 %

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 GBA SimpleStep ELISA® kit is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of GBA protein in human serum, plasma, saliva and cell culture supernatant. Quantitate Human GBA with 10.841 pg/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 (ab315065) 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

GBA also known as glucosylceramidase is a lysosomal enzyme with a molecular mass of approximately 59 kDa. This enzyme breaks down glucosylceramide into glucose and ceramide. GBA is expressed predominantly in tissues with high metabolic activities such as the brain liver and spleen. Its function relies on its catalytic activity where substrates bind to its active site enabling the hydrolysis process necessary for maintaining cellular metabolism.

Biological function summary

GBA plays an important role in sphingolipid metabolism. It participates in the degradation of glycolipids within the lysosome contributing to lipid recycling. It acts independently rather than as a part of a major enzymatic complex. Through its role in degrading glucosylceramide GBA influences cellular homeostasis and bioenergetics ensuring balance in neural and systemic lipid levels.

Pathways

GBA’s enzymatic functions are integral to the glycosphingolipid metabolic pathway. It is involved in the downstream steps of the lysosomal degradation of glycosphingolipids. The pathway operates alongside other important proteins such as beta-glucosidase and CERT-related transfer proteins all of which contribute to membrane lipid organization and signal transduction processes.

Associated diseases and disorders

GBA mutations are linked with Gaucher disease and Parkinson’s disease. In Gaucher disease deficient GBA activity leads to substrate accumulation resulting in hepatosplenomegaly and other systemic symptoms. Reduced GBA activity is also associated with increased alpha-synuclein aggregation in Parkinson’s disease implicating it in the pathogenesis of neurodegenerative disorders. The enzyme’s function in these diseases highlights its role in maintaining cellular equilibrium and signaling pathways.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

  • Sandwich ELISA - Human GBA ELISA Kit (ab315298), expandable thumbnail

    Sandwich ELISA - Human GBA ELISA Kit (ab315298)

    The concentration of GBA was measured in human wild type and knockout HeLa cell extracts based on a 12.5 µg/mL extract load. The mean concentration was determined to be 3,620 pg/mL in wild type extract and undetectable (ND) in knockout extract. The control and GBA knockout cell line extracts are available as Human GBA knockout HeLa cell line ab265038

  • Sandwich ELISA - Human GBA ELISA Kit (ab315298), expandable thumbnail

    Sandwich ELISA - Human GBA ELISA Kit (ab315298)

    Example of human GBA standard curve. Background-subtracted data values (mean +/- SD) are graphed. 

  • Sandwich ELISA - Human GBA ELISA Kit (ab315298), expandable thumbnail

    Sandwich ELISA - Human GBA ELISA Kit (ab315298)

    Interpolated concentration of native GBA was measured in duplicate at different sample concentrations. Undiluted samples are as follows: LNCaP extract 5 µg/mL and kidney extract 50 µg/mL.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in 1X Cell Extraction Buffer PTR. 

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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