Klotho Antibody - BSA Free

Western Blot: Klotho Antibody - BSA Free [NBP1-76511] - Western Blot Validation with Recombinant Protein. Loading: 30 ng of human KLOTHO recombinant protein per lane. Antibodies: KLOTHO NBP1-76511 (Lane 1: 1 ug/mL, Lane 2: 2 ug/mL and Lane 3: 4 ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Immunohistochemistry: Klotho Antibody - BSA Free [NBP1-76511] - Figure 6 Immunohistochemistry Validation of Klotho in Rat Heart Tissue Immunohistochemical analysis of paraffin-embedded rat heart tissue using anti-Klotho antibody at 2.5 u/ml. Tissue was fixed with formaldehyde and blocked with 10% 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.

Immunohistochemistry-Paraffin: Klotho Antibody - BSA Free [NBP1-76511] - Mouse heart tissue diluted at 2.5 ug/ml.

Immunocytochemistry/ Immunofluorescence: Klotho Antibody - BSA Free [NBP1-76511] - Figure 5 Immunofluorescence Validation of Klotho in Mouse Heart TissueImmunofluorescent analysis of 4% paraformaldehyde-fixed mouse heart tissue labeling Klotho with at 20 u/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

Western Blot: Klotho Antibody - BSA Free [NBP1-76511] - Validation in Rat Kidney Tissue LysateLoading: 15ug of lysates per lane.Antibodies: KLOTHO 6107 (1ug/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Western Blot: Klotho Antibody - BSA Free [NBP1-76511] - ARPE-19 cells were divided into the following groups: normal, hypoxia, hypoxia + AAV vector, hypoxia + digoxin, hypoxia + digoxin + nutlin-3a, hypoxia + AAV-p53 mutant infection and hypoxia + miRNA-34a inhibitor. Western blot assay of Klotho in ARPE-19 cells was performed. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33438362/) licensed under a CC-BY license.

Immunohistochemistry: Klotho Antibody - BSA Free [NBP1-76511] - Endothelial klotho staining. Image collected and cropped by CiteAb from the following publication (cardiab.biomedcentral.com/articles/10.1186/s12933-016-0414-5), licensed under a CC-BY license.

Western Blot: Klotho Antibody - BSA Free [NBP1-76511] - Independent Antibody Validation (IAV) via Protein Expression Profile in Human and Mouse Cell Lines. Loading: 15 ug of lysates per lane. Antibodies: KLOTHO NBP1-76511 (2 ug/mL), KLOTHO, competitor antibody (4 ug/mL) and beta-actin (1 ug/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Western Blot: Klotho Antibody - BSA Free [NBP1-76511] - Blockade of the HIF‐1 alpha /p53/miRNA‐34a/Klotho axis decreases the leakage & area of mouse laser‐induced CNV. The mice were divided into the following groups: normal, CNV 7 d, CNV 7 d + 0.1% DMSO, CNV 7 d + digoxin (oral; 2 mg/kg for 7 d), CNV 7 d + AAV‐p53 mutant (intravitreal injection; approximately 3 μL, 3 × 1010 viral particles/mL), CNV 7 d + miRNA‐34a inhibitor (intravitreal injection; 1 μg) & CNV 7 d + AAV‐Klotho full‐length plasmid (intravitreal injection, 2 μL, 5 × 1010 viral particles/mL). A, Western blot was performed to measure HIF‐1 alpha , p‐p53 (S15), p53 (S20), p‐p53 (S46), p53 & Klotho protein levels. B, The relative protein levels of HIF‐1 alpha /GAPDH (B), p‐p53 (S15)/p53 (C), p‐p53 (S20)/p53 (D), p‐p53 (S46)/p53 (E), p53/GAPDH (F) & Klotho/GAPDH (G) were analysed. ***P < .001, CNV 7‐d group vs normal group. ##P < .01, compared with the CNV 7‐d group. NS, CNV 7 d + KL OE group vs CNV 7‐d group. H, RT‐PCR was performed to measure the expression of miRNA‐34a. **P < .01, CNV 7‐d group vs normal group. ##P < .01, compared with the CNV 7‐d group. NS, CNV 7 d + KL OE group vs CNV 7‐d group. I, FFA was performed to measure the leakage of CNV. J, The leakage of CNV was analysed. K, ICGA was performed to measure the area of CNV. L, The area of CNV was analysed. **P < .01, ***P < .001, compared with the CNV 7‐d group Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33438362), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Klotho Antibody - BSA Free [NBP1-76511] - microrna‐34a inhibits the expression of Klotho. ARPE‐19 cells were divided into the following groups: normal, hypoxia, hypoxia + AAV vector, hypoxia + digoxin, hypoxia + digoxin + nutlin‐3a, hypoxia + AAV‐p53 mutant infection & hypoxia + miRNA‐34a inhibitor. A, Western blot assay of Klotho in ARPE‐19 cells was performed. B, The relative protein level of Klotho compared with the GAPDH level was analysed. ***P < .001, hypoxia group vs normal group. ###P < .001, ##P < .01, compared with the hypoxia group. %%P < .01, hypoxia + digoxin + nutlin‐3a group vs hypoxia + digoxin group. C, Schematic of the Klotho 3' UTR with the WT or mutated putative‐binding site of miRNA‐34a inserted into a luciferase (Luc) reporter. D, qRT‐PCR analysis of Klotho mRNA expression in HEK293T cells transfected with miRNA‐34a mimic for 24 h. **P < .01, miRNA‐34a mimic group vs control group. E, Western blot of Klotho in HEK293T cells transfected with a miRNA‐34a mimic for 24 h was performed. F, Immunoblotted Klotho was quantified & normalized to GAPDH. ***P < .001, miRNA‐34a mimic group vs control group. G Relative luciferase activity was detected in HEK293T cells co‐transfected with plasmids containing firefly luciferase & wild‐type (WT) or mutant (Mut) Klotho 3′‐UTR & miRNA‐34a mimic for 24 h. The luciferase activity values were normalized to Renilla reniformis luciferase (TK‐RL) activity. **P < .01, Luc‐Klotho 3′ UTR WT group vs vector group. Statistically non‐significant (NS), Luc‐Klotho 3′ UTR mutant group vs vector group Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/33438362), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Klotho Antibody - BSA Free [NBP1-76511] - High phytate-induced renal phosphate wasting is independent of FGF23.(A) The renal expression of downregulated genes associated with Ca2+ & phosphate homeostasis in rats fed control or phytate-supplemented diets (n = 8 per group). (B) Results of immunoblot analyses & protein levels of renal aKlotho, NHERF1, & NaPi-2a after 2 weeks for rats fed control, HP- LCa2+ , & HP-HCa2+ diets. (C) The levels of renal proteins were quantified using ImageJ software (n = 6 per group). (D, E) Time course analysis of serum levels of intact FGF23 (D) & C-terminal FGF23 (E) in rats fed control, HP-LCa2+ , & HP-HCa2+ diets. (F) Representative immunohistochemical staining of EDTA-decalcified femur sections for FGF23 from rats fed control, HP-LCa2+ , & HP-HCa2+ diets. All data are presented as the mean ± SD of each group (n = 3–8 per group). *, p<0.05; **, p<0.01; ***, p<0.001, compared with controls. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32271147), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt
• Applications: WB, ELISA, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1 mg/ml

Klotho Antibody - BSA Free Summary

• Immunogen: Antibody was raised against a peptide corresponding to 17 amino acids near the center of human Klotho. The immunogen is located within amino acids 410-460 of Klotho. Amino Acid Sequence: SGTTKRRDDAKYMYYLK
• Specificity: Three isoforms of Klotho are known to exist. Human Klotho has 2 isoforms, including isoform 1 (1012aa, 116kD) and isoform 2 (549aa, 62kD). Mouse Klotho has 2 isoforms, including isoform 1 (1014aa, 116kD) and isoform 2 (550aa, 62kD). Rat Klotho has one isoform (1014aa, 117kD). NBP1-76511 can detect human, mouse and rat.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: KL
• Purity: Peptide affinity purified

Applications/Dilutions

• Dilutions:
  • ELISA 1:100-1:2000
  • Immunocytochemistry/ Immunofluorescence 20 ug/mL
  • Immunohistochemistry 2.5 ug/mL
  • Immunohistochemistry-Paraffin 2.5 ug/ml
  • Western Blot 1-4 ug/mL
• Theoretical MW: 116 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control Peptide:

  Klotho Peptide (NBP1-76511PEP)

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Peptide affinity purified

Alternate Names for Klotho Antibody - BSA Free

• EC 3.2.1
• EC 3.2.1.31
• KL
• Klotho

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Klotho Antibody (NBP1-76511)(4)

Publications using NBP1-76511

• Manrique C, Habibi J, Aroor AR et al. Dipeptidyl peptidase-4 inhibition with linagliptin prevents western diet-induced vascular abnormalities in female mice. Cardiovasc Diabetol 2016-07-08 [PMID: 27391040] (ICC/IF)
• Kim OH, Booth CJ, Choi HS et al. High-phytate/low-calcium diet is a risk factor for crystal nephropathies, renal phosphate wasting, and bone loss BioRxiv
• Kamel SS, Baky NAA, Karkeet RM Et al. Astaxanthin extenuates the inhibition of aldehyde dehydrogenase and Klotho protein expression in cyclophosphamide-induced acute cardiomyopathic rat model Clinical and experimental pharmacology & physiology 2021-10-01 [PMID: 34597426] (WB, Rat)
• Kim O H, Booth C J et al. High-phytate/low-calcium diet is a risk factor for crystal nephropathies, renal phosphate wasting, and bone loss. Elife 2020-09-04 [PMID: 32271147] (WB, Rat)

FAQs for Klotho Antibody (NBP1-76511). (Showing 1 - 2 of 2 FAQs).

1. I study physiology (PhD) and I need a klotho kit.
   • A list of our klotho products can be found here. I am not entirely sure what you are looking for as I don't have your complete message for some reason. Can you please clarify what you are looking for?
2. This data sheet contains figures including a western blot and fluorescence images of rat or mouse heart tissue. Can you give me the information about the source of these figures?
   • The images supplied on the datasheet for NBP1-76511 were generated from our lab's testing of this antibody.

Bioinformatics

• Gene Symbol: KL
• Uniprot:
  • NP_004786(Human)