WB
Western blot analysis of TIM 3 on different lysates with Rabbit anti-TIM 3 antibody 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) Exposure time: 40 seconds; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of RPMI 8226 (positive) and HT-29 (negative) labeling TIM 3 with Rabbit anti-TIM 3 antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
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 (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
TIM 3 was immunoprecipitated from 0.2 mg RPMI 8226 cell lysate with abbit anti-TIM 3 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using abbit anti-TIM 3 antibody at 1/2,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: RPMI 8226 cell lysate (input), Lane 2: abbit anti-TIM 3 antibody IP in RPMI 8226 cell lysate, Lane 3: Rabbit IgG instead of abbit anti-TIM 3 antibody in RPMI 8226 cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 59 seconds.| Product Name | TIM 3 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human TIM 3 aa 1-223. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIPWB |
| WB:1:2000 ICC:1:100 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | HAVCR2 |
|---|---|
| Gene Synonyms | TIM3 CD366 KIM-3 SPTCL TIMD3 Tim-3 TIMD-3 HAVcr-2 |
| Gene Full Name | hepatitis A virus cellular receptor 2 |
| Gene Summary | The protein encoded by this gene belongs to the immunoglobulin superfamily, and TIM family of proteins. CD4-positive T helper lymphocytes can be divided into types 1 (Th1) and 2 (Th2) on the basis of their cytokine secretion patterns. Th1 cells are involved in cell-mediated immunity to intracellular pathogens and delayed-type hypersensitivity reactions, whereas, Th2 cells are involved in the control of extracellular helminthic infections and the promotion of atopic and allergic diseases. This protein is a Th1-specific cell surface protein that regulates macrophage activation, and inhibits Th1-mediated auto- and alloimmune responses, and promotes immunological tolerance. [provided by RefSeq, Sep 2011] |
| Alternative Names | CD366 antibody FLJ14428 antibody HAVcr-2 antibody Havcr2 antibody HAVR2_HUMAN antibody Hepatitis A virus cellular receptor 2 antibody Kidney injury molecule 3 antibody KIM 3 antibody KIM3 antibody T cell immunoglobulin and mucin domain containing 3 antibody CD366 antibody FLJ14428 antibody HAVcr-2 antibody Havcr2 antibody HAVR2_HUMAN antibody Hepatitis A virus cellular receptor 2 antibody Kidney injury molecule 3 antibody KIM 3 antibody KIM3 antibody T cell immunoglobulin and mucin domain containing 3 antibody T cell immunoglobulin mucin 3 antibody T-cell immunoglobulin and mucin domain-containing protein 3 antibody T-cell immunoglobulin mucin family member 3 antibody T-cell immunoglobulin mucin receptor 3 antibody T-cell membrane protein 3 antibody Tim 3 antibody TIM-3 antibody TIM3 antibody TIMD-3 antibody TIMD3 antibody |
| Molecular Weight(MW) | 33kDa(Observed band size: 55kDa) |
| Cellular Localization | Membrane, Cell junction, Cell membrane. |
WB
Western blot analysis of TIM 3 on different lysates with Rabbit anti-TIM 3 antibody 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) Exposure time: 40 seconds; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of RPMI 8226 (positive) and HT-29 (negative) labeling TIM 3 with Rabbit anti-TIM 3 antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
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 (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
TIM 3 was immunoprecipitated from 0.2 mg RPMI 8226 cell lysate with abbit anti-TIM 3 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using abbit anti-TIM 3 antibody at 1/2,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: RPMI 8226 cell lysate (input), Lane 2: abbit anti-TIM 3 antibody IP in RPMI 8226 cell lysate, Lane 3: Rabbit IgG instead of abbit anti-TIM 3 antibody in RPMI 8226 cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 59 seconds.| Application Notes | WB:1:2000 ICC:1:100 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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. |
