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    products/cell-lysates/human-uchl1-pgp95-knockout-hek-293t-cell-lysate-ab263773.pdf

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Human UCHL1 (PGP9.5) knockout HEK-293T cell lysate (ab263773)

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Western blot - Human UCHL1 knockout HEK293T cell lysate (ab263773)
  • Sanger Sequencing - Human UCHL1 knockout HEK293T cell lysate (ab263773)

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Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
Knockout
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Human UCHL1 (PGP9.5) knockout HEK-293T cell line (ab255443)
Protein
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Recombinant human PGP9.5 protein (ab53962)

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Overview

  • Product name

    Human UCHL1 (PGP9.5) knockout HEK-293T cell lysate
    See all PGP9.5 kits
  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    HEK293T
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous: 45 bp deletion in exon 1.
  • Passage number

    <20
  • Knockout validation

    Sanger Sequencing, Western Blot (WB)
  • Reconstitution notes

    To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.

    *Usage of SDS sample buffer is not recommended with these lyophilized lysates.

  • Notes

    Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.

    User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

    Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
    See here for more information on knockout cell lysates.

    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.

    This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab255547 - Human UCHL1 knockout HEK293T cell lysate 1 x 100µg
    ab255594 - Human wild-type HEK293T cell lysate 1 x 100µg
  • Research areas

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Parkinson's disease
    • Parkin / PARK
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Soma marker
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Deubiquitination
    • Tags & Cell Markers
    • Cell Type Markers
    • Neuroscience Markers
    • Neuronal
    • Neuroscience
    • Diseases
  • Cell type

    epithelial
  • STR Analysis

    Amelogenin X D5S818: 8, 9 D13S317: 11, 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 15, 20 TH01: 7, 9.3 TPOX: 11, 12 CSF1PO: 12

Target

  • 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.
  • Target information above from: UniProt accession P09936 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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

Associated products

  • KO cell lines

    • Human UCHL1 (PGP9.5) knockout HEK-293T cell line (ab255443)
  • Related Products

    • Anti-PGP9.5 antibody [EPR4118] - Neuronal Marker (ab108986)
    • Anti-PGP9.5 antibody [EPR4118] - BSA and Azide free (ab220823)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab263773 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
WB
Use at an assay dependent concentration.
Notes
WB
Use at an assay dependent concentration.

Images

  • Western blot - Human UCHL1 knockout HEK293T cell lysate (ab263773)
    Western blot - Human UCHL1 knockout HEK293T cell lysate (ab263773)

    Lane 1: Wild-type Hap1 cell lysate (20 µg)

    Lane 2: UCHL1 knockout Hap1 cell lysate (20 µg)

    Lane 3: SH-SY5Y cell lysate (20 µg)

    Lane 4: Wild-type HEK-293T cell lysate (20 µg)

    Lane 5: UCHL1 knockout HEK-293T cell lysate (20 µg)

    Lanes 1 - 5: Merged signal (red and green). Green - ab108986 observed at 25 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab108986 was shown to react with PGP9.5 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab255443 (knockout cell lysate ab263773) was used. Wild-type and PGP9.5 knockout samples were subjected to SDS-PAGE. ab108986 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively.Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Sanger Sequencing - Human UCHL1 knockout HEK293T cell lysate (ab263773)
    Sanger Sequencing - Human UCHL1 knockout HEK293T cell lysate (ab263773)
    Homozygous: 45 bp deletion in exon 1

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab263773? Please let us know so that we can cite the reference in this datasheet.

ab263773 has not yet been referenced specifically in any publications.

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