Overview

  • Product name

    Human UCHL1 ELISA Kit
    See all PGP9.5 kits
  • Detection method

    Colorimetric
  • Precision

    Inter-assay
    Sample n Mean SD CV%
    293T extract 3 5.1%
  • Sample type

    Cell culture supernatant, Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    48 pg/ml
  • Range

    313 pg/ml - 20000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 117 108% - 123%
    Tissue Extracts 114 109% - 119%
    Cell culture media 97 96% - 98%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Product overview

    UCHL1 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of UCHL1 protein in human cell culture extracts, tissue extracts and cell culture supernatant.


    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 substrate 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.


    Ubiquitin carboxy-terminal hydrolase L1 (UCHL1), also known as PGP9.5, is a deubiquitinating thiol protease that hydrolyzes at the C-terminal glycine of ubiquitin, resulting in the ubiquitin monomer. Although mature UCHL1 is comprised only of 220 amino acids, it forms complicated knot structures, which is thought to protect the protein from unintended proteasomal degradation. It is present in all neurons, cells of the neuroendocrine system, associated tumors, and in the testes/ovaries. UCHL1 is highly abundant in neurons, comprising 1-2% of total brain protein. The UCHL1 gene is required for normal synaptic and cognitive function. Polymorphisms or loss of gene function have been associated with Parkinson’s and Alzheimer’s diseases.


    This kit is knockout validated for specifically detecting UCHL1 in human HAP1 cells.


    Serum and plasma samples are not compatible with this kit due to severe matrix effects.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human UCHL1 Capture Antibody 1 x 600µl
    10X Human UCHL1 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml
    Human UCHL1 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • Function

    Ubiquitin-protein hydrolase involved both in the processing of ubiquitin precursors and of ubiquitinated proteins. This enzyme is a thiol protease that recognizes and hydrolyzes a peptide bond at the C-terminal glycine of ubiquitin. Also binds to free monoubiquitin and may prevent its degradation in lysosomes. The homodimer may have ATP-independent ubiquitin ligase activity.
  • Tissue specificity

    Found in neuronal cell bodies and processes throughout the neocortex (at protein level). Expressed in neurons and cells of the diffuse neuroendocrine system and their tumors. Weakly expressed in ovary. Down-regulated in brains from Parkinson disease and Alzheimer disease patients.
  • Involvement in disease

    Parkinson disease 5
    Neurodegeneration with optic atrophy, childhood-onset
  • Sequence similarities

    Belongs to the peptidase C12 family.
  • Post-translational
    modifications

    O-glycosylated.
  • Cellular localization

    Cytoplasm. Endoplasmic reticulum membrane. About 30% of total UCHL1 is associated with membranes in brain.
  • Information by UniProt
  • Alternative names

    • Epididymis luminal protein 117
    • Epididymis secretory protein Li 53
    • HEL 117
    • HEL S 53
    • NDGOA
    • Neuron cytoplasmic protein 9.5
    • OTTHUMP00000218137
    • OTTHUMP00000218139
    • OTTHUMP00000218140
    • OTTHUMP00000218141
    • Park 5
    • PARK5
    • PGP 9.5
    • PGP9.5
    • PGP95
    • Protein gene product 9.5
    • Ubiquitin C terminal esterase L1
    • Ubiquitin C terminal hydrolase
    • Ubiquitin C terminal hydrolase L1
    • Ubiquitin carboxyl terminal esterase L1
    • Ubiquitin carboxyl terminal hydrolase isozyme L1
    • Ubiquitin carboxyl-terminal hydrolase isozyme L1
    • Ubiquitin thioesterase L1
    • Ubiquitin thiolesterase
    • Ubiquitin thiolesterase L1
    • UCH-L1
    • UCHL1
    • UCHL1_HUMAN
    see all
  • Database links

Associated products

Applications

Our Abpromise guarantee covers the use of ab229201 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

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • The UCHL1 standard curve was prepared as described in Section 10. Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of UCHL1 were measured in duplicates, interpolated from the UCHL1 standard curves and corrected for sample dilution. Undiluted samples are as follows: human brain extract 2.5 µg/mL, A549 extract 50 µg/mL, SH-SY5Y extract 2.5 µg/mL, and 293T extract 15 µg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean UCHL1 concentration in the undiluted samples was determined to be 7.5 ng/mL in brain extract, 15 ng/mL in A549 extract, 5.5 ng/mL in SH-SY5Y extract, and 5.2 ng/mL in 293T extract.

  • The concentrations of UCHL1 were measured in duplicate and interpolated from the UCHL1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • The concentrations of UCHL1 were measured in duplicates, interpolated from the UCHL1 standard curves and corrected for sample dilution. Undiluted samples are as follows: mouse brain extract 2.5 µg/mL and rat brain extract 1.25 µg/mL. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean UCHL1 concentration in the undiluted samples was determined to be 3.7 ng/mL in mouse brain extract and 2.9 ng/mL in rat brain extract.

  • Raw data is plotted for wild-type and UCHL1-knockout HAP1 cell extracts tested at 18 μg/mL.

Protocols

References

ab229201 has not yet been referenced specifically in any publications.

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