Recombinant
RabMAb

Recombinant Anti-Klotho antibody [EPR6856] - BSA and Azide free (ab232366)

Overview

  • Product name

    Anti-Klotho antibody [EPR6856] - BSA and Azide free
    See all Klotho primary antibodies
  • Description

    Rabbit monoclonal [EPR6856] to Klotho - BSA and Azide free
  • 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: WB, IHC-Pmore details
  • 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

    • WB: Wild Type Mouse kidney tissue lysate.
  • General notes

    Ab232366 is the carrier-free version of ab181373. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab232366 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab232366 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. Detects a band of approximately 62 kDa (predicted molecular weight: 62, 116 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181373).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181373).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181373).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181373).

References

ab232366 has not yet been referenced specifically in any publications.

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