Recombinant
RabMAb

Recombinant Anti-Klotho antibody [EPR6856] (ab181373)

Rabbit recombinant monoclonal Klotho antibody [EPR6856]. Validated in WB, IHC and tested in Mouse, Human. Cited in 6 publication(s).

Overview

  • Product name

    Anti-Klotho antibody [EPR6856]
    See all Klotho primary antibodies
  • Description

    Rabbit monoclonal [EPR6856] to Klotho
  • Host species

    Rabbit
  • Specificity

    When we tested the antibody in human samples, the only form of the protein that we are able to detect is the secreted one (about 65kDa). When we tested the antibody in mouse samples (wt vs KO) we could only detect the membrane bound form of the protein (about 130kDa). This difference could be due to the nature of the samples (different species). However, we do not have a clear explanation about this. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented here.
  • Tested applications

    Suitable for: IHC-P, WBmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human Klotho aa 400-500. The exact sequence is proprietary.
    Database link: Q9UEF7

  • Positive control

    • Human testis, fetal kidney and fetal thymus lysates; Human clear cell carcinoma kidney tissue, Human kidney tissue.
  • General notes

     

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab181373 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
IHC-P 1/300. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

For unpurified, use 1/100 - 1/250.

WB 1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 62, 116 kDa).
  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    • Function

      May have weak glycosidase activity towards glucuronylated steroids. However, it lacks essential active site Glu residues at positions 239 and 872, suggesting it may be inactive as a glycosidase in vivo. May be involved in the regulation of calcium and phosphorus homeostasis by inhibiting the synthesis of active vitamin D (By similarity). Essential factor for the specific interaction between FGF23 and FGFR1.
      The Klotho peptide generated by cleavage of the membrane-bound isoform may be an anti-aging circulating hormone which would extend life span by inhibiting insulin/IGF1 signaling.
    • Tissue specificity

      Present in cortical renal tubules (at protein level). Soluble peptide is present in serum and cerebrospinal fluid. Expressed in kidney, placenta, small intestine and prostate. Down-regulated in renal cell carcinomas, hepatocellular carcinomas, and in chronic renal failure kidney.
    • Involvement in disease

      Defects in KL are a cause of hyperphosphatemic familial tumoral calcinosis (HFTC) [MIM:211900]. A severe metabolic disorder that manifests with hyperphosphatemia and massive calcium deposits in the skin and subcutaneous tissues. Some patients manifest recurrent, transient, painful swellings of the long bones associated with the radiographic findings of periosteal reaction and cortical hyperostosis and absence of skin involvement.
    • Sequence similarities

      Belongs to the glycosyl hydrolase 1 family. Klotho subfamily.
    • Domain

      Contains 2 glycosyl hydrolase 1 regions. However, the first region lacks the essential Glu active site residue at position 239, and the second one lacks the essential Glu active site residue at position 872.
    • Post-translational
      modifications

      N-glycosylated.
    • Cellular localization

      Secreted and Cell membrane. Isoform 1 shedding leads to a soluble peptide.
    • Information by UniProt
    • Database links

    • Alternative names

      • Kl antibody
      • KLOT_HUMAN antibody
      • Klotho peptide antibody

    Images

    • All lanes : Anti-Klotho antibody [EPR6856] (ab181373) at 5 µg/ml

      Lane 1 : Wild Type (WT) Mouse Kidney Tissue Lysate
      Lane 2 : Knock Out (KO) Mouse Kidney Tissue Lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 62, 116 kDa



      The blots were produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane were blocked for an hour before being incubated with ab181373  (rabbit anti-Klotho antibody; diluted 5 ug/ml) for 24 hours at 4°C. Antibody binding was detected using infrared labelled goat anti-rabbit (green) antibody at 1:10,000 dilutions for 1 hour at room temperature before imaging.

    • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Klotho with unpurified ab181373 at 1/100 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • All lanes : Anti-Klotho antibody [EPR6856] (ab181373) at 1/1000 dilution (purified)

      Lane 1 : Human fetal kidney
      Lane 2 : Human testis
      Lane 3 : Human fetal thymus

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : HRP anti-rabbit IgG, specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 62, 116 kDa
      Observed band size: 62 kDa
      why is the actual band size different from the predicted?



      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunohistochemical staining of paraffin embedded human kidney with purified ab181373 at a working dilution of 1 in 300. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    • All lanes : Anti-Klotho antibody [EPR6856] (ab181373) at 1/1000 dilution (unpurified)

      Lane 1 : Human fetal kidney lysates
      Lane 2 : Human testis lysates
      Lane 3 : Human fetal thymus lysates

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 62, 116 kDa

    • Immunohistochemical analysis of paraffin-embedded human kidney clear cell carcinoma tissue labeling Klotho with unpurified ab181373 at 1/100 dilution.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    References

    This product has been referenced in:

    • Li D  et al. Enhanced Expression of Secreted a-Klotho in the Hippocampus Alters Nesting Behavior and Memory Formation in Mice. Front Cell Neurosci 13:133 (2019). Read more (PubMed: 31001090) »
    • Smith ER  et al. aKlotho-FGF23 interactions and their role in kidney disease: a molecular insight. Cell Mol Life Sci N/A:N/A (2019). Read more (PubMed: 31350618) »
    See all 9 Publications for this product

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