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<strong>Figure 1 Western Blot Validation in Human Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: TIM-1 3809 (8 μg/mL), overnight incubation at 4° C in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: TIM-1 3809 (8 μg/mL), TIM-1 3811 (1 μg/mL), beta-actin (1 μg/mL), and GAPDH (0.02 μg/mL), overnight incubation at 4° C (3809) or 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 3 Validation with TIM-1 siRNA Knockdown in Hela Cells</strong><br>HeLa cells were transfected with control siRNAs (lane 1) or TIM-1 siRNAs (lane 2) Loading: 15 μg of HeLa whole cell lysates per lane.Antibodies: 3809 (8 μg/mL), 1 h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Western Blot Validation in Mouse Tissues </strong><br>Loading: 15 μg of lysates.Antibodies: TIM-1 3809, 2 μ g/mL, 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 5 Immunofluorescence Validation of TIM-1 in Human Testis</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed human testis tissue labeling TIM-1 with 3809 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).
<strong>Figure 6 Immunofluorescence Validation of TIM-1 in Mouse Kidney</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed mouse kidney tissue labeling TIM-1 with 3809 at 10 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red) and DAPI staining (blue).
Figure 1 Western Blot Validation in Human Cell Lines
Loading: 15 μg of lysates per lane.Antibodies: TIM-1 3809 (8 μg/mL), overnight incubation at 4° C in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
IAV
KD
Trial Size

TIM-1 Antibody

CATALOG NUMBER: 3809

Clonality:
Polyclonal
Tested Applications:
ELISA, WB, IHC-P/IF
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human, Mouse
Homology:
Predicted species reactivity based on immunogen sequence: Rat: (56.3%)
Immunogen:
Anti-TIM-1 antibody (3809) was raised against a peptide corresponding to 16 amino acids near the amino terminus of human TIM-1.
The immunogen is located within amino acids 50 - 100 of TIM-1.
Conjugate:
Unconjugated
Tested Applications:
ELISA, WB, IHC-P/IF
Application Note:
WB: 1 - 8 μg/mL (overnight incubation at 4° C); IHC-P: 10 μg/mL; IF: 20 μg/mL.
Antibody validated: Western Blot in human and mouse samples; Immunohistochemistry and Immunofluorescence in human samples. All other applications and species not yet tested.
Positive Control 1:
Cat. No. 1202 - A431 Cell Lysate
Positive Control 2:
Cat. No. 1203 - A549 Cell Lysate
Positive Control 3:
Cat. No. 1224 - Daudi Cell Lysate
Positive Control 4:
Cat. No. 11-401 - Human Uterus Tissue Slide
Predicted Molecular Weight:
Predicted: 44kD
Observed: 60kD (Post-modification: 4 N-linked glycosylation)
Validation:

Independent Antibody Validation in Cell lines (Figure 2) shows similar TIM-1 expression profile in human cell lines detected by two independent anti-TIM-1 antibodies that recognize different epitopes, 3809 against N-terminus domain and 3811 against the central domain. TIM-1 proteins are detected in all the tested cell lines except MCF-7 at different expression levels by the two independent antibodies. 

siRNA knockdown validation (Figure 3): Anti-TIM-1 antibody (3809) specificity was further verified by TIM-1 specific siRNA knockdown. TIM-1 signal in HeLa cells transfected with TIM-1 siRNAs was much weaker in comparison with that in HeLa cells transfected with control siRNAs.

Isoforms:
Human TIM-1 has 5 isoforms, including isoform a precursor (364aa, 39kD), isoform a (364aa, 39kD), isoform b precursor (401aa, 44kD), isoform b (359aa, 39kD), and isoform c (359aa, 39kD). This antibody detects human isoform b precursor. Mouse TIM-1 has 3 isoforms, including isoform a precursor (305aa, 33kD), isoform b precursor (282aa, 31kD) and isoform b (282aa, 31kD). Rat TIM-1 has only one isoform identified so far (307aa, 34kD).
Purification:
TIM-1 Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
physical-state:
Liquid
Buffer:
TIM-1 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
TIM-1 antibody can be stored at 4°C up to one year. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
HAVCR1
Additional Names:
TIM-1 Antibody: TIM, KIM1, TIM1, HAVCR, KIM-1, TIM-1, TIMD1, TIMD-1, HAVCR-1, Hepatitis A virus cellular receptor 1, Kidney injury molecule 1, HAVcr-1
Protein Accession Number:
NP_036338
Protein Gi Number:
153085427
Ncbi Gene Id Number:
26762
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
TIM-1 Antibody: The human form of TIM-1 was initially discovered as a membrane glycoprotein through which the hepatitis A virus can gain entry into a cell. It was also identified as kidney injury molecule 1 (Kim-1), a predicted adhesion molecule that is upregulated on the surfaces of kidney epithelia. It is also expressed on T helper 2 (Th2) cells of the immune system, and following the binding of its natural ligand TIM-4, stimulates T cell expansion and cytokine production in response to viral challenge. It has been suggested that hyperactivation of TIM-1 leads to an increased level of Th2 responsiveness and asthma susceptibility, and antibodies to TIM-1 may therefore be a novel approach to treating asthma.
Background Reference 1:
Feigelstock et al. J. Virol. 1998; 72:6621-8
Background Reference 2:
Ichimura et al. J. Biol. Chem. 1998; 273:4135-42
Background Reference 3:
Meyers et al. Mol. Med. 2005; 11:362-9
Background Reference 4:
Mayers et al. Nat. Immunol. 2005;6:455-64.

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:

3809

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

Datasheet
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