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Human ANPEP (CD13) knockout THP-1 cell lysate (ab275505)

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Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (ab275505)
  • Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (ab275505)
  • Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (ab275505)
  • Sanger Sequencing - Human ANPEP knockout THP-1 cell lysate (ab275505)

Overview

  • Product name

    Human ANPEP (CD13) knockout THP-1 cell lysate
    See all CD13 kits

  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    THP-1

  • Organism

    Human

  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 2

  • 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

Target

  • Function

    Broad specificity aminopeptidase. Plays a role in the final digestion of peptides generated from hydrolysis of proteins by gastric and pancreatic proteases. May play a critical role in the pathogenesis of cholesterol gallstone disease. May be involved in the metabolism of regulatory peptides of diverse cell types including small intestinal and tubular epithelial cells, macrophages, granulocytes and synaptic membranes from the CNS. Found to cleave antigen peptides bound to major histocompatibility complex class II molecules of presenting cells and to degrade neurotransmitters at synaptic junctions. Is also implicated as a regulator of IL-8 bioavailability in the endometrium, and therefore may contribute to the regulation of angiogenesis. Is used as a marker for acute myeloid leukemia and plays a role in tumor invasion. In case of human coronavirus 229E (HCoV-229E) infection, serves as receptor for HCoV-229E spike glycoprotein. Mediates as well human cytomegalovirus (HCMV) infection.

  • Tissue specificity

    Expressed in epithelial cells of the kidney, intestine, and respiratory tract; granulocytes, monocytes, fibroblasts, endothelial cells, cerebral pericytes at the blood-brain barrier, synaptic membranes of cells in the CNS. Also expressed in endometrial stromal cells, but not in the endometrial glandular cells. Found in the vasculature of tissues that undergo angiogenesis and in malignant gliomas and lymph node metastases from multiple tumor types but not in blood vessels of normal tissues. A soluble form has been found in plasma. It is found to be elevated in plasma and effusions of cancer patients.

  • Sequence similarities

    Belongs to the peptidase M1 family.

  • Domain

    Amino acids 260-353 are essential to mediate susceptibility to infection with HCoV-229E (in porcine/human chimeric studies) and more specifically amino acids 288-295 (mutagenesis studies).

  • Post-translational
    modifications

    Sulfated.
    N- and O-glycosylated.
    May undergo proteolysis and give rise to a soluble form.

  • Cellular localization

    Cell membrane. Cytoplasm > cytosol. A soluble form has also been detected.
  • Information by UniProt

  • Alternative names

    • Alanyl (membrane) aminopeptidase
    • Alanyl aminopeptidase
    • Aminopeptidase M
    • Aminopeptidase N
    • AMPN_HUMAN
    • ANPEP
    • AP M
    • AP N
    • AP-M
    • AP-N
    • APN
    • CD 13
    • CD13
    • CD13 antigen
    • gp150
    • hAPN
    • LAP 1
    • LAP1
    • Microsomal aminopeptidase
    • Myeloid plasma membrane glycoprotein CD13
    • p150
    • PEPN
    see all

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab275505 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. Predicted molecular weight: 109 kDa.
Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 109 kDa.

Images

  • Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (ab275505)
    Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (ab275505)

    Lane 1: Wild-type THP-1 cell lysate 30 ug
    Lane 2: ANPEP knockout THP-1 cell lysate 30 ug
    Lane 3: PANC-1 cell lysate 30 ug
    Lane 4: HEK-293 cell lysate 30 ug
    Lanes 1 - 4:Merged signal (red and green). Green - ab227663 observed at 160 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
    ab227663 was shown to react with CD13 in wild-type THP-1 cells in western blot with loss of signal observed in ANPEP knockout cell line ab273759 (knockout cell lysate ab275505). Wild-type and CD13 knockout THP-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab227663 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 400 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 hour at room temperature before imaging.

  • Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (ab275505)
    Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (ab275505)

    Lane 1: Wild-type THP-1 cell lysate 30 ug
    Lane 2: ANPEP knockout THP-1 cell lysate 30 ug
    Lane 3: PANC-1 cell lysate 30 ug
    Lane 4: HEK-293 cell lysate 30 ug
    Lanes 1 - 4:Merged signal (red and green). Green - ab108382 observed at 160 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
    ab108382 was shown to react with CD13 in wild-type THP-1 cells in western blot with loss of signal observed in ANPEP knockout cell line ab273759 (knockout cell lysate ab275505). Wild-type and CD13 knockout THP-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab108382 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 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 hour at room temperature before imaging.

  • Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (ab275505)
    Western blot - Human ANPEP (CD13) knockout THP-1 cell lysate (ab275505)

    Lane 1: Wild-type THP-1 cell lysate 30 ug
    Lane 2: ANPEP knockout THP-1 cell lysate 30 ug
    Lane 3: PANC-1 cell lysate 30 ug
    Lane 4: HEK-293 cell lysate 30 ug
    Lanes 1 - 4:Merged signal (red and green). Green - ab108310 observed at 160 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
    ab108310 was shown to react with CD13 in wild-type THP-1 cells in western blot with loss of signal observed in ANPEP knockout cell line ab273759 (knockout cell lysate ab275505). Wild-type and CD13 knockout THP-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab108310 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 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 hour at room temperature before imaging.

  • Sanger Sequencing - Human ANPEP knockout THP-1 cell lysate (ab275505)
    Sanger Sequencing - Human ANPEP knockout THP-1 cell lysate (ab275505)

    Homozygous: 2 bp deletion in exon 2

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

References (0)

Publishing research using ab275505? Please let us know so that we can cite the reference in this datasheet.

ab275505 has not yet been referenced specifically in any publications.

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