Invitrogen

Klotho Polyclonal Antibody

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Klotho Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of paraffin-embedded mouse heart tissue using Klotho Polyclonal Antibody (Product # PA5-21078) at 2.5 µg/mL. Tissue was fixed with formaldehyde and blocked with 0.1 serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Klotho Antibody in Western Blot (WB) — Western Blot Validation with Recombinant Protein. Loading: 30 ng of human KLOTHO recombinant protein per lane. Antibodies: Klotho Polyclonal Antibody (Product # PA5-21078) (Lane 1: 1 µg/mL, Lane 2: 2 µg/mL and Lane 3: 4 µg/mL), 1h incubation at RT in 0.05 NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10,000 dilution. Observed at around 72 kDa.

Klotho Antibody in Immunocytochemistry (ICC/IF) — Fig 3 miR-200c decreases KLOTHO expression in HK-2 cells by translational repression. (A) KLOTHO protein expression in HK-2 cells 24 hrs after transfection with 25 nM mimic control or miR-200c mimic. Cells were cultured for another 48 hrs without mimic control or miR-200c mimic before sampling. Band intensities were analyzed and normalized against TUBA using densitometry. (B) A KLOTHO 3'-UTR reporter plasmid in combination with 50 nM mimic control or miR-200c mimic was transfected into HK-2 cells for 4 hrs. After a medium change, HK-2 cells were cultured for another 12 hrs before sampling. Luciferase activity was normalized against protein amount. (C) HK-2 cells were transfected with 50 nM mimic control or miR-200c mimic for 4 hrs and cultured for another 24 hrs before harvesting total RNA. KLOTHO mRNA levels were evaluated by q-PCR. (D) Mutations were introduced into the 3'-UTR of KLOHO mRNA as indicated. (E) The effect of 100 nM miR-200c mimic on the reporter activity of wild type (WT, pMirTarget-KL3'-UTR-WT) and mutant (MUT, pMirTarget-KL3'-UTR-MUT) plasmids in HK-2 cells was measured by luciferase assay. (F) HK-2 cells were stained with anti-KLOTHO antibody and Alexa Fluor 488-labeled goat anti-rabbit IgG. KLOTHO protein was evaluated under fluorescence microscopy. Scale bar = 10 mum. Values represent individual measurements and the mean +- SD. Data were analyzed using the Mann-Whitney U -test or the Mann-Whitney U -test with Bonferroni correction. * P < 0.05, n

Klotho Antibody in Western Blot, Neutralization (WB, Neu) — Fig. 5 FGF23 attenuates SDF-1-induced CXCR4 expression via interaction of FGF receptor-1 (FGFR1), but not Klotho. A FGFR-1, -2, -3, or -4 mRNA expression levels in EPCs were detected by RT-PCR. HK-2 lysates were used as positive controls. B EPCs were pre-treated with FGF23 and/or FGFR inhibitor (PD173074) for 30 min followed by SDF-1 stimulation for 15 min or 6 h. C EPCs harboring control or Klotho shRNA were pre-treated with FGF23 for 30 min followed by SDF-1 stimulation for 10 min or 6 h. D EPCs were pre-treated with Klotho neutralizing antibody for 60 min and FGF23 for 30 min followed by SDF-1 stimulation for 10 min or 24 h. E EPCs were treated with different doses of Klotho. F EPCs were collected at the indicated time points after Klotho treatment. G EPCs harboring control or Klotho shRNA were pre-treated with FGF23 for 24 h. Protein lysates were analyzed by immunoblot using indicated antibodies. beta-actin or GAPDH were used as protein loading controls. The numbers under the gel lanes represent the relative protein level.

Product Details

PA5-21078

Applications	Tested Dilution	Publications
Western Blot (WB)	1-2 µg/mL	View 4 publications 4 publications
Immunohistochemistry (IHC)	2.5 µg/mL	-
Immunocytochemistry (ICC/IF)	-	View 1 publication 1 publication
Neutralization (Neu)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Human, Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	A 17 amino acid peptide near the center of human KLOTHO.
Conjugate	Unconjugated
Form	Liquid
Concentration	1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS
Contains	0.02% sodium azide
Storage conditions	Maintain refrigerated at 2-8°C for up to 3 months. For long term storage store at -20°C
RRID	AB_11153007

Product Specific Information

A suggested positive control is HepG2 cell lysate.

PA5-21078 can be used with blocking peptide PEP-1192.

Target Information

KLOTHO is the systemic anti-aging hormone within the glycosidase1 superfamily. It encodes a type I membrane protein that is abundant in the kidney and brain. In mice, a deficiency in KLOTHO expression leads to various systemic phenotypes resembling human aging such as arteriosclerosis, osteoporosis, and skin atrophy together with growth retardation, short life-span and infertility. Transgenic mice overexpressing KLOTHO have an extended life span by inhibiting insulin/IGF1 signaling. KLOTHO is involved in the regulation of calcium/phosphorus homeostasis by inhibiting the synthesis of active vitamin D and identified as a potential tumor suppressor.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

FGF23 ameliorates ischemia-reperfusion induced acute kidney injury via modulation of endothelial progenitor cells: targeting SDF-1/CXCR4 signaling.

Cell death & disease

Chang HM,Peng KY,Chan CK,Sun CY,Chen YY,Chang HM,Huang CL,Liu PC,Chen PY,Wang KC,Wang WJ,Wu CC,Lin YF,Lai TS,Huang TM,Young GH,Lin SL,Ostermann M,Chu TS,Chueh JS,Wu VC

PA5-21078 was used in Western Blot to highlight a revolutionary and important role that FGF23 plays in the nephroprotection of IR-AKI.

Sat Apr 17 00:00:00 EDT 2021

FGF23 ameliorates ischemia-reperfusion induced acute kidney injury via modulation of endothelial progenitor cells: targeting SDF-1/CXCR4 signaling.

Cell death & disease

Chang HM,Peng KY,Chan CK,Sun CY,Chen YY,Chang HM,Huang CL,Liu PC,Chen PY,Wang KC,Wang WJ,Wu CC,Lin YF,Lai TS,Huang TM,Young GH,Lin SL,Ostermann M,Chu TS,Chueh JS,Wu VC

PA5-21078 was used in Western Blot to highlight a revolutionary and important role that FGF23 plays in the nephroprotection of IR-AKI.

Sat Apr 17 00:00:00 EDT 2021

FGF23 ameliorates ischemia-reperfusion induced acute kidney injury via modulation of endothelial progenitor cells: targeting SDF-1/CXCR4 signaling.

Cell death & disease

Chang HM,Peng KY,Chan CK,Sun CY,Chen YY,Chang HM,Huang CL,Liu PC,Chen PY,Wang KC,Wang WJ,Wu CC,Lin YF,Lai TS,Huang TM,Young GH,Lin SL,Ostermann M,Chu TS,Chueh JS,Wu VC

PA5-21078 was used in Western Blot to highlight a revolutionary and important role that FGF23 plays in the nephroprotection of IR-AKI.

Sat Apr 17 00:00:00 EDT 2021

microRNA-200c regulates KLOTHO expression in human kidney cells under oxidative stress.

PloS one

Morii K,Yamasaki S,Doi S,Irifuku T,Sasaki K,Doi T,Nakashima A,Arihiro K,Masaki T

PA5-21078 was used in Immunocytochemistry-immunoflourescence to study KLOTHO expression and its regulation by miR-200c.

Fri Feb 14 00:00:00 EST 2020

Cells

Mytych J,Sołek P,Będzińska A,Rusinek K,Warzybok A,Tabęcka-Łonczyńska A,Koziorowski M

PA5-21078 was used in Western Blotting to provide evidence of klotho involvement in the crosstalk on the line ER-Golgi, which may, in turn, affect activation of SASP.

Tue Jan 21 00:00:00 EST 2020

Cells

Mytych J,Sołek P,Będzińska A,Rusinek K,Warzybok A,Tabęcka-Łonczyńska A,Koziorowski M

PA5-21078 was used in Western Blotting to provide evidence of klotho involvement in the crosstalk on the line ER-Golgi, which may, in turn, affect activation of SASP.

Tue Jan 21 00:00:00 EST 2020

The Role of the Anti-Aging Protein Klotho in IGF-1 Signaling and Reticular Calcium Leak: Impact on the Chemosensitivity of Dedifferentiated Liposarcomas.

Cancers

Delcroix V,Mauduit O,Tessier N,Montillaud A,Lesluyes T,Ducret T,Chibon F,Van Coppenolle F,Ducreux S,Vacher P

PA5-21078 was used in Western Blotting to provide a new therapeutic strategy in the management of DDLPS.

Wed Nov 14 00:00:00 EST 2018

Protective role of klotho protein on epithelial cells upon co-culture with activated or senescent monocytes.

Experimental cell research

Mytych J,Wos I,Solek P,Koziorowski M

PA521078 was used in immunocytochemistry and western blot to show that ROS-induced DNA damage reduces metabolic activity, inhibits proliferation, and induces cell cycle arrest followed by activation of p16-, p21-, and p27- mediated DNA damage response pathways, premature senescence, and apoptosis induction

Sun Jan 15 00:00:00 EST 2017

Bioinformatics

Protein Aliases: HFTC3; Klotho; Klotho peptide; secreted form of Klotho protein

Gene Aliases: alpha-kl; KL

UniProt ID: (Human) Q9UEF7, (Rat) Q9Z2Y9, (Mouse) O35082

Entrez Gene ID: (Human) 9365, (Rat) 83504, (Mouse) 16591

Molecular Function: glycosidase

Function(s)

MAPK cascade carbohydrate metabolic process energy reserve metabolic process aging insulin receptor signaling pathway fibroblast growth factor receptor signaling pathway regulation of phosphatidylinositol 3-kinase signaling positive regulation of bone mineralization phosphatidylinositol-3-phosphate biosynthetic process positive regulation of GTPase activity phosphatidylinositol phosphorylation phosphatidylinositol-mediated signaling calcium ion homeostasis positive regulation of MAPKKK cascade by fibroblast growth factor receptor signaling pathway glycosyl compound metabolic process acute inflammatory response metabolic process

Process(es)

beta-glucuronidase activity signal transducer activity Ras guanyl-nucleotide exchange factor activity fibroblast growth factor receptor binding hormone activity vitamin D binding beta-glucosidase activity 1-phosphatidylinositol-3-kinase activity fibroblast growth factor binding phosphatidylinositol-4,5-bisphosphate 3-kinase activity hydrolase activity, hydrolyzing O-glycosyl compounds protein binding hydrolase activity hydrolase activity, acting on glycosyl bonds

Klotho Polyclonal Antibody

Product Details

PA5-21078

Western Blot (WB)

Immunohistochemistry (IHC)

Immunocytochemistry (ICC/IF)

Neutralization (Neu)

Species Reactivity

Published species

Host/Isotype

Class

Type

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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