TIM 3 Recombinant Rabbit Monoclonal Antibody [PSH10-00]
cat.: HA723169
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH10-00 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 33 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human TIM 3 aa 1-223. |
| Positive control: | RPMI 8226 cell lysate, HUT 102 cell lysate, RPMI 8226. |
| Subcellular location: | Membrane, Cell junction, Cell membrane. |
| Recommended Dilutions:
WB IF-Cell FC IP |
1:2,000 1:100 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: Q8TDQ0 Human |
| Alternative names: | CD366 FLJ14428 HAVcr-2 Havcr2 HAVR2_HUMAN Hepatitis A virus cellular receptor 2 Kidney injury molecule 3 KIM 3 KIM3 T cell immunoglobulin and mucin domain containing 3 T cell immunoglobulin mucin 3 T-cell immunoglobulin and mucin domain-containing protein 3 T-cell immunoglobulin mucin family member 3 T-cell immunoglobulin mucin receptor 3 T-cell membrane protein 3 Tim 3 TIM-3 TIM3 TIMD-3 TIMD3 |
Images
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Fig1:
Western blot analysis of TIM 3 on different lysates with Rabbit anti-TIM 3 antibody (HA723169) at 1/2,000 dilution. Lane 1: RPMI 8226 cell lysate (20 µg/Lane) Lane 2: HT-29 cell lysate (negative) (20 µg/Lane) Lane 3: HUT 102 cell lysate (20 µg/Lane) Predicted band size: 33 kDa Observed band size: 55 kDa Exposure time: 40 seconds; ECL: K1802; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723169) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of RPMI 8226 (positive) and HT-29 (negative) labeling TIM 3 with Rabbit anti-TIM 3 antibody (HA723169) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TIM 3 antibody (HA723169) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Flow cytometric analysis of HT-29 (left, negative) and RPMI 8226 (right, positive) cells labeling TIM 3. Cells were fixed and permeabilized. Then stained with the primary antibody (HA723169, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig4:
TIM 3 was immunoprecipitated from 0.2 mg RPMI 8226 cell lysate with HA723169 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723169 at 1/2,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: RPMI 8226 cell lysate (input) Lane 2: HA723169 IP in RPMI 8226 cell lysate Lane 3: Rabbit IgG instead of HA723169 in RPMI 8226 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 59 seconds; ECL: K1802 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.