Overview

  • Product name
    Human Clusterin ELISA Kit 
    See all Clusterin kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Serum 9 2.7%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 4.9%
  • Sample type
    Cell culture supernatant, Saliva, Milk, Urine, Serum, Cell culture extracts, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    22.5 pg/ml
  • Range
    234.4 pg/ml - 15000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 103 100% - 106%
    Saliva 100 99% - 100%
    Milk 99 97% - 101%
    Urine 102 101% - 102%
    Serum 101 98% - 103%
    Cell culture extracts 105 96% - 112%
    Heparin Plasma 102 101% - 103%
    EDTA Plasma 101 99% - 102%
    Citrate Plasma 107 106% - 109%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human, Rhesus monkey
    Does not react with: Mouse, Rat, Cow
  • Product overview

    Clusterin in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Clusterin protein in human serum, plasma, urine, saliva, milk, cell culture supernatant, and cell extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB Development Solution is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Clusterin (CLU) is composed of an antiparallel, disulfide-linked heterodimer of an alpha chain and a beta chain that self-associate and can form higher oligomers. It is ubiquitously expressed in various tissues. Clusterin functions as an extracellular chaperone that prevents aggregation of nonnative proteins. It does not require ATP and does not refold proteins by itself. Clusterin does so by maintaining partially unfolded proteins in a state appropriate for subsequent refolding by other chaperones, such as HSPA8/HSC70. Clusterin prevents stress-induced aggregation of blood plasma proteins by inhibiting the formation of amyloid fibrils through APP, APOC2, B2M, CALCA, CSN3, SNCA and aggregation-prone LYZ variants (in vitro). It can bind to cell surface receptors and trigger internalization of the chaperone-client complex and subsequent lysosomal or proteasomal degradation.


    Sensitivity:


    Samples diluted in Sample Diluent NS -  36.1 pg/mL 


    Samples diluted in 1X Cell Extraction Buffer PTR - 22.5 pg/mL   

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

Applications

Our Abpromise guarantee covers the use of ab174447 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • The Clusterin standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • The Clusterin standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Clusterin were measured in duplicates, interpolated from the Clusterin standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:80,000, plasma (heparin) 1:40,000, plasma (EDTA) 1:40,000, and plasma (citrate) 1:20,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Clusterin concentration in the neat samples was determined to be 174 µg/mL in serum, 123 µg/mL in plasma (heparin), 158 µg/mL in plasma (EDTA), and 206 µg/mL in plasma (citrate).

  • The concentrations of Clusterin were measured in duplicates, interpolated from the Clusterin standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 1:32, saliva 1:80, and milk 1:3,200. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Clusterin concentration in the neat samples was determined to be 157 ng/mL in urine, 246 ng/mL in saliva, and 20,323 ng/mL in milk.

  • The concentrations of Clusterin were measured in duplicates, interpolated from the Clusterin standard curves and corrected for sample dilution. Undiluted samples are as follows: HepG2 supernatant 1:12, HepG2 extract 200 µg/mL, and MDA-MB-231 extract 750 µg/mL.

  • The concentrations of Clusterin were measured in duplicate, interpolated from the Clusterin standard curve and corrected for sample dilution. Undiluted samples are 1:20,000. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Clusterin concentration in the neat sample was determined to be 94 µg/mL.

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Clusterin concentration was determined to be 150 µg/mL with a range of 92 – 227 µg/mL.

  • Standard curve comparison between human Clusterin SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

  • Standard curve comparison between current human FGF19 SimpleStep ELISA® kit and original human FGF19 SimpleStep ELISA® kit. The current SimpleStep ELISA kit shows increased sensitivity.

Protocols

References

ab174447 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Easy to use and reliable.

Excellent Excellent 5/5 (Ease of Use)
Abreviews
This kit was easy to use and reliable. The aasay is easily performed within a 4 hour time frame and worked very well with EDTA-plasma samples diluted into the range 1:10,000 - 1: 20,000 with good reproducibility between duplicates.

I would thoroughly recommend this product.
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Verified customer

Submitted Apr 18 2015

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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