Recombinant Anti-Klotho antibody [EPR6856] (ab181373)
![Western blot - Anti-Klotho antibody [EPR6856] (ab181373)](https://www.abcam.com/ps/products/181/ab181373/Images/ab181373-256508-anti-klotho-antibody-epr6856-western-blot.jpg "Western blot - Anti-Klotho antibody [EPR6856] (ab181373)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6856] to Klotho
- Suitable for: IHC-P, WB
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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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.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6856 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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) |
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).
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| Notes |
|---|
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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. |
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WB
1/1000. Detects a band of approximately 62 kDa (predicted molecular weight: 62, 116 kDa). |
Target
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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
modificationsN-glycosylated. -
Cellular localization
Secreted and Cell membrane. Isoform 1 shedding leads to a soluble peptide. - Information by UniProt
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Database links
- Entrez Gene: 9365 Human
- Entrez Gene: 16591 Mouse
- Omim: 604824 Human
- SwissProt: Q9UEF7 Human
- SwissProt: O35082 Mouse
- Unigene: 524953 Human
- Unigene: 6500 Mouse
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Alternative names
- Kl antibody
- KLOT_HUMAN antibody
- Klotho peptide antibody
Images
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Western blot - Anti-Klotho antibody [EPR6856] (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 kDaThe 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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Klotho antibody [EPR6856] (ab181373)](https://www.abcam.com/ps/products/181/ab181373/Images/ab181373-210753-anti-klotho-antibody-epr6856-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Klotho antibody [EPR6856] (ab181373)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.
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![Western blot - Anti-Klotho antibody [EPR6856] (ab181373)](https://www.abcam.com/ps/products/181/ab181373/Images/ab181373-239394-anti-klotho-antibody-epr6856-western-blot.jpg)
Western blot - Anti-Klotho antibody [EPR6856] (ab181373)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
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Klotho antibody [EPR6856] (ab181373)](https://www.abcam.com/ps/products/181/ab181373/Images/ab181373-239395-anti-klotho-antibody-epr6856-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Klotho antibody [EPR6856] (ab181373)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.
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![Western blot - Anti-Klotho antibody [EPR6856] (ab181373)](https://www.abcam.com/ps/products/181/ab181373/Images/ab181373-210755-anti-klotho-antibody-epr6856-western-blot.jpg)
Western blot - Anti-Klotho antibody [EPR6856] (ab181373)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 -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Klotho antibody [EPR6856] (ab181373)](https://www.abcam.com/ps/products/181/ab181373/Images/ab181373-210754-anti-klotho-antibody-epr6856-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Klotho antibody [EPR6856] (ab181373)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.
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![Anti-Klotho antibody [EPR6856] (ab181373)](https://www.abcam.com/ps/products/181/ab181373/Images/ab181373-7-benefits-of-recombinant-antibodies.png)
Anti-Klotho antibody [EPR6856] (ab181373)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (30)
ab181373 has been referenced in 30 publications.
- Li S et al. Abnormally decreased renal Klotho is linked to endoplasmic reticulum-associated degradation in mice. Int J Med Sci 19:321-330 (2022). PubMed: 35165517
- Xiong G et al. Long non-coding RNA MEG3 regulates the progress of osteoarthritis by regulating the miR-34a/Klotho axis. Ann Transl Med 10:454 (2022). PubMed: 35571440
- Li C et al. Comprehensive Profiling Reveals Distinct Microenvironment and Metabolism Characterization of Lung Adenocarcinoma. Front Genet 12:619821 (2021). PubMed: 34122500
- Zhu L et al. Klotho inhibits H2 O2 -induced oxidative stress and apoptosis in periodontal ligament stem cells by regulating UCP2 expression. Clin Exp Pharmacol Physiol 48:1412-1420 (2021). PubMed: 34174105
- Li M et al. METTL14 promotes glomerular endothelial cell injury and diabetic nephropathy via m6A modification of α-klotho. Mol Med 27:106 (2021). PubMed: 34503454