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Human CSTB (Cystatin-B) knockout HEK-293T cell lysate (ab257905)

Overview

  • Product name

    Human CSTB (Cystatin-B) knockout HEK-293T cell lysate

  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    HEK293T

  • Organism

    Human

  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous: Insertion of the selection cassette in exon 1.

  • Passage number

  • Knockout validation

    Sanger Sequencing

  • 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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

  • Tested applications

    Suitable for: WBmore details

Properties

Target

  • Function

    This is an intracellular thiol proteinase inhibitor. Tightly binding reversible inhibitor of cathepsins L, H and B.

  • Involvement in disease

    Defects in CSTB are the cause of progressive myoclonic epilepsy type 1 (EPM1) [MIM:254800]. EPM1 is an autosomal recessive disorder characterized by severe, stimulus-sensitive myoclonus and tonic-clonic seizures. The onset, occurring between 6 and 13 years of age, is characterized by convulsions. Myoclonus begins 1 to 5 years later. The twitchings occur predominantly in the proximal muscles of the extremities and are bilaterally symmetrical, although asynchronous. At first small, they become late in the clinical course so violent that the victim is thrown to the floor. Mental deterioration and eventually dementia develop.

  • Sequence similarities

    Belongs to the cystatin family.

  • Cellular localization

    Cytoplasm. Nucleus.
  • Information by UniProt

  • Alternative names

    • CHROW21
    • CPI B
    • CPI-B
    • CST 6
    • CST6
    • CSTB
    • Cystatin B
    • Cystatin B (stefin B)
    • Cystatin-B
    • CYTB
    • CYTB_HUMAN
    • EPM1
    • EPM1A
    • Liver thiol proteinase inhibitor
    • PME
    • Stefin-B
    • STF B
    • STFB
    • ULD
    see all

Properties

Images

  • Western blot - Human CSTB (Cystatin-B) knockout HEK293T cell lysate (ab257905)
    Western blot - Human CSTB (Cystatin-B) knockout HEK293T cell lysate (ab257905)

    Lane 1:Wild-type HEK293T cell lysate (20 ug)
    Lane 2:CSTB knockout HEK293T cell lysate (20 ug)
    Lane 3:U87-MG cell lysate (20 ug)
    Lane 4:JAR cell lysate (20 ug)

    ab92449 was shown to specifically react with Cystatin-B in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266321 (knockout cell lysate ab257905) was used. Wild-type and Cystatin-B knockout samples were subjected to SDS-PAGE. ab92449 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 CSTB knockout HEK293T cell lysate (ab257905)
    Sanger Sequencing - Human CSTB knockout HEK293T cell lysate (ab257905)
    Homozygous: Insertion of the selection cassette in exon 1

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