Anti-KLOTHO antibody was prepared from whole rabbit serum produced by repeated immunizations with a 16 amino acid synthetic peptide near the internal region of human KLOTHO.
Anti-KLOTHO Antibody has been tested for use in ELISA, Western Blotting, Immunohistochemistry and Immunofluorescence. Specific conditions for reactivity should be optimized by the end user. Expect a band at approximately 116 kDa in Western Blots of speci
Immunofluorescence Validation of KLOTHO. Tissue: Mouse Heart Tissue. Fixation: 4% paraformaldehyde-fixed. Primary Antibody: KLOTHO with at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1:500 dilution (red).
Immunohistochemistry Validation of KLOTHO. Tissue: Rat Heart Tissue. Fixation: paraffin-embedded, formaldehyde and blocked with 10% serum for 1 h at RT. Antigen retrieval: heat mediation with a citrate buffer (pH6). Primary Antibody: anti-KLOTHO antibody at 2.5 µg/ml overnight at 4C. Secondary Antibody: goat anti-rabbit IgG H&L (HRP) at 1:250. Counter stained with Hematoxylin.
Western Blot Validation. Load: 15 µg of human or mouse lysates per lane. Lane 1: 293, Lane 2: A431, Lane 3: A549, Lane 4: CaCo-2, Lane 5: HepG2, Lane 6: K562, Lane 7: MCF7, Lane 8: Jurkat, Lane 9: SK-N-SH, Lane 10: THP-1, Lane 11: NIH/3T3. Primary Antibody: KLOTHO at 2µg/mL, competitor antibody at 4 µg/mL, and beta-actin at 1 µg/mL for 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Immunohistochemistry Validation of KLOTHO. Tissue: Mouse Heart Tissue. Fixation: paraffin-embedded, formaldehyde and blocked with 10% serum for 1 h at RT. Antigen retrieval: heat mediation with a citrate buffer (pH6). Primary Antibody: anti-KLOTHO antibody at 2.5 µg/ml overnight at 4C. Secondary Antibody: goat anti-rabbit IgG H&L (HRP) at 1:250. Counter stained with Hematoxylin.
Western Blot Validation. Load: 30 ng of human KLOTHO recombinant protein per lane. Primary Antibody: KLOTHO at (Lane 1: 1 µg/mL, Lane 2: 2 µg/mL and Lane 3: 4 µg/mL), for 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution. Observed at around 72kD.
Western Blot Validation. Load: 15 µg of rat kidney lysate. Primary Antibody: KLOTHO at 1 µg/mL for 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
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