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<strong>Figure 1 Independent Antibody Validation (IAV) via Protein Expression Profile in Human and Mouse Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: KLOTHO 6107 (2 μg/mL), KLOTHO, competitor antibody (4 μg/mL) and beta-actin 3779 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Western Blot Validation with Recombinant Protein</strong><br>Loading: 30 ng of human KLOTHO recombinant protein per lane.Antibodies: KLOTHO 6107 (Lane 1: 1 μg/mL, Lane 2: 2 μg/mL and Lane 3: 4 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.Observed at around 72kD.
<strong>Figure 3 Western Blot Validation in Rat Kidney Tissue Lysate</strong><br>Loading: 15 μg of lysates per lane.Antibodies: KLOTHO 6107 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunohistochemistry Validation of KLOTHO in Mouse Heart Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded mouse heart tissue using anti-KLOTHO antibody (6107) at 2.5 μg/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.
<strong>Figure 5 Immunofluorescence Validation of KLOTHO in Mouse Heart Tissue</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse heart tissue labeling KLOTHO with 6107 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
<strong>Figure 6 Immunohistochemistry Validation of KLOTHO in Rat Heart Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded rat heart tissue using anti-KLOTHO antibody (6107) at 2.5 μg/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.
Figure 1 Independent Antibody Validation (IAV) via Protein Expression Profile in Human and Mouse Cell Lines
Loading: 15 μg of lysates per lane.Antibodies: KLOTHO 6107 (2 μg/mL), KLOTHO, competitor antibody (4 μg/mL) and beta-actin 3779 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
Trial Size

KLOTHO Antibody

CATALOG NUMBER: 6107

Clonality:
Polyclonal
Tested Applications:
ELISA, IF, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse, Rat
Immunogen:
Anti-KLOTHO antibody (6107) 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.
Conjugate:
Unconjugated
Tested Applications:
ELISA, IF, IHC-P, WB
Application Note:
WB: 1-4 μg/mL; IHC: 2.5 μg/mL; IF: 20 μg/mL.
Antibody validated: Western Blot in human, mouse and rat samples; Immunohistochemistry in mouse and rat samples; Immunofluorescence in mouse samples. All other applications and species not yet tested.
Specificity:
Three isoforms of KLOTHO are known to exist.
Positive Control 1:
Cat. No. 1203 - A549 Cell Lysate
Positive Control 2:
Cat. No. 1204 - K562 Cell Lysate
Positive Control 3:
Cat. No. 1205 - Jurkat Cell Lysate
Positive Control 4:
Cat. No. 1282 - 3T3/NIH Cell Lysate
Predicted Molecular Weight:
Predicted: 116kD
Observed: 116kD
Validation:

Independent Antibody Validation in Cell lines (Figure 1) shows similar KLOTHO expression profile in human and mouse cell lines detected by two independent anti-KLOTHO antibodies that recognize different epitopes, 6107 against central domain and competitor antibody against human recombinant protein.  KLOTHO proteins are detected in the most tested cell lines at different expression levels by the two independent antibodies. 

Recombinant Protein Test (Figure 2): Anti-KLOTHO antibodies (6107) detected human KLOTHO recombinant protein at different concentrations.

Isoforms:
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). 6107 can detect human, mouse and rat.
Purification:
KLOTHO Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
KLOTHO Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
KLOTHO antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
KL
Additional Names:
KLOTHO Antibody: Klotho
Protein Accession Number:
NP_004786
Protein Gi Number:
24497614
Ncbi Gene Id Number:
9365
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
KLOTHO Antibody: 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.
Background Reference 1:
Kuro-o et al. Nature1997; 390:45-51.
Background Reference 2:
Kurosu et al. Science2005; 309:1829-33.
Background Reference 3:
Liu et al. Science 2007; 317:803-6.
Background Reference 4:
Wolf et al. Oncogene2008; 27:7094-105.

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

6107

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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