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Human CANX (Calnexin) knockout HEK-293T cell lysate (ab263805)

Overview

  • Product name

    Human CANX (Calnexin) knockout HEK-293T cell lysate
    See all Calnexin kits

  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    HEK293T

  • Organism

    Human

  • Mutation description

    Knockout achieved by using CRISPR/Cas9, 19 bp deletion in exon2.

  • 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 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
    ab255473 - Human CANX knockout HEK293T cell lysate 1 x 100µg
    ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg

  • Research areas

  • Cell type

    epithelial

  • STR Analysis

    Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12

Target

  • Function

    Calcium-binding protein that interacts with newly synthesized glycoproteins in the endoplasmic reticulum. It may act in assisting protein assembly and/or in the retention within the ER of unassembled protein subunits. It seems to play a major role in the quality control apparatus of the ER by the retention of incorrectly folded proteins.

  • Sequence similarities

    Belongs to the calreticulin family.

  • Cellular localization

    Endoplasmic reticulum membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt

  • Alternative names

    • Calnexin
    • CALX_HUMAN
    • CANX
    • CNX
    • FLJ26570
    • Histocompatibility complex class I antigen binding protein p88
    • IP90
    • Major histocompatibility complex class I antigen-binding protein p88
    • p90
    see all

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab255473 - Human CANX knockout HEK293T cell lysate 1 x 100µg
    ab255553 - Human wild-type HEK293T cell lysate 1 x 100µg

  • Research areas

  • Cell type

    epithelial

  • STR Analysis

    Amelogenin X D5S818: 8, 9 D13S317: 12, 14 D7S820: 11 D16S539: 9, 13 vWA: 16, 19 TH01: 7, 9.3 TPOX: 11 CSF1PO: 11, 12

Images

  • Western blot - Human CANX knockout HEK293T cell lysate (ab263805)
    Western blot - Human CANX knockout HEK293T cell lysate (ab263805)

    Lane 1: Wild-type HEK-293T cell lysate (20 ug)
    Lane 2:     CANX knockout HEK-293T cell lysate (20 ug)
    Lane 3:     U-2 OS cell lysate (20 ug)
    Lane 4:     MCF7 cell lysate (20 ug)

    Lanes 1 - 4: Merged signal (red and green). Green - ab92573 observed at 80 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

    ab92573 was shown to react with Calnexin in wild-type HEK-293T cells in Western blot with loss of signal observed in CANX knockout cell line ab255368 (CANX knockout cell lysate ab263805). Wild-type HEK-293T and CANX knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab92573 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 20000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Western blot - Human CANX (Calnexin) knockout HEK293T cell lysate (ab263805)
    Western blot - Human CANX (Calnexin) knockout HEK293T cell lysate (ab263805)

    Lane 1: wild-type HEK-293T cell lysate 20 ug
    Lane 2: CANX knockout HEK-293T cell lysate 20 ug
    Lane 3: U-2 OS cell lysate 20 ug
    Lane 4: MCF7 cell lysate 20 ug
    Lanes 1 - 4:Merged signal (red and green). Green - ab238078 observed at 80 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
    ab238078 was shown to react with Calnexin in wild-type HEK-293T cells in Western blot with loss of signal observed in CANX knockout cell line ab255368 (CANX knockout cell lysate ab263805). Wild-type HEK-293T and CANX knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab238078 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at 1 ug/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Western blot - Human CANX (Calnexin) knockout HEK293T cell lysate (ab263805)
    Western blot - Human CANX (Calnexin) knockout HEK293T cell lysate (ab263805)
    Lane 1: Wild-type HEK-293T cell lysate (20µg)

    Lane 2: CANX knockout HEK-293T cell lysate (20µg)
    Lanes 1- 2: Merged signal (red and green). Green - ab133615 observed at 90 kDa. Red - loading control ab8245 observed at 37 kDa.
    ab133615 Recombinant Anti-Calnexin antibody [EPR3632] was shown to specifically react with CANX in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255368 (knockout cell lysate ab263805) was used. Wild-type and CANX knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133615 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 CANX knockout HEK293T cell lysate (ab263805)
    Sanger Sequencing - Human CANX knockout HEK293T cell lysate (ab263805)
    Homozygous: 19 bp deletion in exon2

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