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Human COL4A4 (Collagen alpha-4(IV) chain) knockout HeLa cell lysate (ab258824)

Overview

  • Product name

    Human COL4A4 (Collagen alpha-4(IV) chain) knockout HeLa cell lysate

  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    HeLa

  • Organism

    Human

  • Mutation description

    Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon29 and 2 bp insertion in exon29.

  • 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.

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab263038 - Human COL4A4 knockout HeLa cell lysate 1 x 100µg
    ab255929 - Human wild-type HeLa cell lysate 1 x 100µg

  • Cell type

    epithelial

  • Disease

    Adenocarcinoma

  • Gender

    Female

  • STR Analysis

    Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10

Target

  • Function

    Type IV collagen is the major structural component of glomerular basement membranes (GBM), forming a 'chicken-wire' meshwork together with laminins, proteoglycans and entactin/nidogen.

  • Tissue specificity

    Alpha 3 and alpha 4 type IV collagens are colocalized and present in kidney, eye, basement membranes of lens capsule, cochlea, lung, skeletal muscle, aorta, synaptic fibers, fetal kidney and fetal lung. PubMed:8083201 reports similar levels of expression of alpha 3 and alpha 4 type IV collagens in kidney, but PubMed:7523402 reports that in kidney levels of alpha 3 type IV collagen are significantly lower than those of alpha 4 type IV collagen. Highest levels of expression of alpha 4 type IV collagen are detected in kidney, calvaria, neuroretina and cardiac muscle. Lower levels of expression are observed in brain, lung and thymus, and no expression is detected in choroid plexus, liver, adrenal, pancreas, ileum or skin.

  • Involvement in disease

    Alport syndrome, autosomal recessive
    Hematuria, benign familial

  • Sequence similarities

    Belongs to the type IV collagen family.
    Contains 1 collagen IV NC1 (C-terminal non-collagenous) domain.

  • Domain

    Alpha chains of type IV collagen have a non-collagenous domain (NC1) at their C-terminus, frequent interruptions of the G-X-Y repeats in the long central triple-helical domain (which may cause flexibility in the triple helix), and a short N-terminal triple-helical 7S domain.

  • Post-translational
    modifications

    Prolines at the third position of the tripeptide repeating unit (G-X-Y) are hydroxylated in some or all of the chains.
    Type IV collagens contain numerous cysteine residues which are involved in inter- and intramolecular disulfide bonding. 12 of these, located in the NC1 domain, are conserved in all known type IV collagens.
    The trimeric structure of the NC1 domains is stabilized by covalent bonds between Lys and Met residues.

  • Cellular localization

    Secreted > extracellular space > extracellular matrix > basement membrane. Colocalizes with COL4A4 and COL4A5 in GBM, tubular basement membrane (TBM) and synaptic basal lamina (BL).
  • Information by UniProt

  • Alternative names

    • CO4A4_HUMAN
    • COL4A4
    • Collagen alpha-4(IV) chain

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab263038 - Human COL4A4 knockout HeLa cell lysate 1 x 100µg
    ab255929 - Human wild-type HeLa cell lysate 1 x 100µg

  • Cell type

    epithelial

  • Disease

    Adenocarcinoma

  • Gender

    Female

  • STR Analysis

    Amelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10

Images

  • Sanger Sequencing - Human COL4A4 knockout HeLa cell lysate (ab258824)
    Sanger Sequencing - Human COL4A4 knockout HeLa cell lysate (ab258824)

    Allele-1: 1 bp deletion in exon29

     

  • Sanger Sequencing - Human COL4A4 knockout HeLa cell lysate (ab258824)
    Sanger Sequencing - Human COL4A4 knockout HeLa cell lysate (ab258824)

    Allele-2: 2 bp insertion in exon29

     

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