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Human ERP29 knockout HEK-293T cell lysate (ab257188)

Overview

  • Product name

    Human ERP29 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: 1 bp insertion in exon 1.

  • Passage number

  • 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, 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

  • Relevance

    Proper protein folding and post-translational modifications are essential for secretory protein export out of the endoplasmic reticulum. This task is accomplished by chaperone proteins such as protein disulfide isomerase (PDI), GRP94, and BiP. A recently characterized protein, designated ERp29, is closely related to these chaperone proteins and appears to be upregulated during ER stress conditions. ERp29 is a soluble 259-residue protein that is localized to the lumen of the endoplasmic reticulum in all mammalian cells. Research has shown that there are two primary domains within ERp29. The first is the C-terminal region that is a novel, all helical, fold that is most likely involved with ERp29 retention to the ER. The second is the N-terminal region that resembles that of PDI’s thioredoxin module. The protein shows sequence similarity to the protein disulfide isomerase family. However, it lacks the thioredoxin motif characteristic of this family, suggesting that this protein does not function as a disulfide isomerase. The protein dimerizes and is thought to play a role in the processing of secretory proteins within the ER.

  • Cellular localization

    Endoplasmic reticulum, Cell surface.

  • Alternative names

    • C12orf8
    • Chromosome 12 open reading frame 8
    • Endoplasmic reticulum lumenal protein ERp28
    • Endoplasmic reticulum protein 29
    • Endoplasmic reticulum protein 29 isoform 1
    • Endoplasmic reticulum protein ERp29
    • Endoplasmic reticulum resident protein 28
    • Endoplasmic reticulum resident protein 29
    • Endoplasmic reticulum resident protein 31
    • Epididymis secretory protein Li 107
    • ERp28
    • ERP29_HUMAN
    • ERp31
    • HEL S 107
    • PDI-DB
    • PDIA9
    • Protein disulfide isomerase family A member 9
    see all

Properties

Images

  • Western blot - Human ERP29 knockout HEK293T cell lysate (ab257188)
    Western blot - Human ERP29 knockout HEK293T cell lysate (ab257188)

    Lane 1: Wild-type HEK293T cell lysate (20 µg)

    Lane 2: ERP29 knockout HEK293T cell lysate (20 µg)

    Lane 3: HepG2 cell lysate (20 µg)

    Lanes 1-3: Merged signal (red and green). Green - ab176573 observed at 29 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab176573 Anti-ERp29 antibody [EPR12499(B)] was shown to specifically react with ERp29 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266543 (knockout cell lysate ab257188) was used. Wild-type and ERp29 knockout samples were subjected to SDS-PAGE. ab176573 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 10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Human ERP29 knockout HEK293T cell lysate (ab257188)
    Western blot - Human ERP29 knockout HEK293T cell lysate (ab257188)

    Lane 1: Wild-type HEK293T cell lysate (20 µg)

    Lane 2: ERP29 knockout HEK293T cell lysate (20 µg)

    Lane 3: HepG2 cell lysate (20 µg)

    Lanes 1-3: Merged signal (red and green). Green - ab175193 observed at 29 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab175193 Anti-ERp29 antibody [EPR12985] was shown to specifically react with ERp29 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266543 (knockout cell lysate ab257188) was used. Wild-type and ERp29 knockout samples were subjected to SDS-PAGE. ab175193 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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 10000 dilution for 1 hour at room temperature before imaging.

  • Sanger Sequencing - Human ERP29 knockout HEK293T cell lysate (ab257188)
    Sanger Sequencing - Human ERP29 knockout HEK293T cell lysate (ab257188)
    Homozygous: 1 bp insertion in exon 1

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