Revision 8

#45208

Store at -20C

TIM-3 (D5D5R™) XP® Rabbit mAb
CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IP, IHC-Bond, IHC-P, FC-L

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
45-70

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q8TDQ0

Entrez-Gene Id:
84868

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:50 - 1:200
Immunohistochemistry (Paraffin) 1:200 - 1:800
Flow Cytometry (Live) 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #81229.

Specificity/Sensitivity

TIM-3 (D5D5R™) XP® Rabbit mAb recognizes endogenous levels of total TIM-3 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the extracellular domain of human TIM-3 protein.

Background

T cell Ig- and mucin-domain-containing molecules (TIMs) are a family of transmembrane proteins expressed by various immune cells. TIM-3 is an inhibitory molecule that is induced following T cell activation (1-3 ). TIM-3 is expressed by exhausted T cells in the settings of chronic infection and cancer (4,5), and tumor-infiltrating T cells that coexpress PD-1 and TIM-3 exhibit the most severe exhausted phenotype (5). Tumor-infiltrating dendritic cells (DCs) also express TIM-3. TIM-3 expression on DCs was found to suppress innate immunity by reducing the immunogenicity of nucleic acids released by dying tumor cells (6). Research studies show that heterodimerization of TIM-3 with CEACAM-1 is critical for the inhibitory function of TIM-3, and co-blockade of TIM-3 and CEACAM-1 enhanced anti-tumor responses in a mouse model of colorectal cancer (7). In addition, blockade of TIM-3 in mouse models of autoimmunity enhanced the severity of disease (1). Finally, binding of Galectin-9 to TIM-3 expressed by Th1 cells induces T cell death (8).

Background References

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) FC-L: Flow Cytometry (Live)

Cross-Reactivity Key

H: Human

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意，否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件，除非书 CST 的合法授书代表以书面形式书独接受， 否书均被拒书，并且无效。

专品专有“专供研究使用”的专专或专似的专专声明， 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的， 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专，且专用于研专用途。将专品用于专断、专防或治专目的， 或专专售（专独或作专专成）或其他商专目的而专专专品，均需要 CST 的专独专可。客专：(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品，或使用专品制造任何商专专品，(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专，或使用专品开专任何与 CST 的专品或服专专争的专品或服专， (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专，(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 ， (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 8

#45208

TIM-3 (D5D5R™) XP® Rabbit mAb

Cell Signaling Technology Logo
Western blot analysis of extracts from primary human CD4+ T cells and various cell lines using TIM-3 (D5D5R™) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). CD4+ T cells were purified from human blood and stimulated for 9 days using beads coated with CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (6.7 ng/ml).
Western Blotting Image 1: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Western blot analysis of extracts from various cell lines using TIM-3 (D5D5R™) XP® Rabbit mAb #45208 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 2: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Western blot analysis of extracts from 293T cells, untransfected (-) or transfected with a construct expressing full-length human Myc/DDK-tagged TIM-3 (hTIM-3-Myc/DDK; +), using TIM-3 (D5D5R™) XP® Rabbit mAb.
Western Blotting Image 3: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 8

#45208

TIM-3 (D5D5R™) XP® Rabbit mAb

Cell Signaling Technology Logo
Immunoprecipitation of TIM-3 from RPMI 8226 cell extracts. Lane 1 is 10% input, lane 2 is precipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TIM-3 (D5D5R™) XP® Rabbit mAb, #45208. Western blot was performed using TIM-3 (D5D5R™) XP® Rabbit mAb. Secondary detection was performed using #12291, Protein A (HRP Conjugate).
Immunoprecipitation Image 1: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded renal clear cell carcinoma using TIM-3 (D5D5R™) XP® Rabbit mAb performed on the Leica® BOND™ Rx.
Immunohistochemistry Image 1: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human tonsil using TIM-3 (D5D5R™) XP® Rabbit mAb.
Immunohistochemistry Image 1: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 8

#45208

TIM-3 (D5D5R™) XP® Rabbit mAb

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using TIM-3 (D5D5R™) XP® Rabbit mAb.
Immunohistochemistry Image 2: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using TIM-3 (D5D5R™) XP® Rabbit mAb. Note staining of alveolar macrophages.
Immunohistochemistry Image 3: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Immunohistochemical analysis of paraffin-embedded cell pellets, primary CD4+ T cells (left) and HT-29 cells (right), using TIM-3 (D5D5R™) XP® Rabbit mAb. CD4+ T cells were purified from human blood and stimulated for 7 days using beads coated with CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (6.7 ng/ml).
Immunohistochemistry Image 4: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 8

#45208

TIM-3 (D5D5R™) XP® Rabbit mAb

Cell Signaling Technology Logo
Multiplex immunohistochemical analysis of paraffin-embedded human breast carcinoma usng TIM-3 (D5D5R™) XP® rabbit mAb (yellow), PD-1 (D4W2J) XP® rabbit mAb #86163 (green), PD-L1 (E1L3N®) XP® rabbit mAb #13684 (red), LAG3 (D2G4O™) XP® rabbit mAb #15372 (magenta), CD8α (C8/144B) mouse mAb #70306 (orange), and Pan-keratin (C11) mouse mAb #4545 (cyan).
Immunohistochemistry Image 5: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Multiplex immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using TIM-3 (D5D5R™) XP® rabbit mAb (yellow), PD-1 (EH33) mouse mAb #43248 (green), CD8α (C8/144B) mouse mAb #70306 (magenta), CD68 (D4B9C) XP® rabbit mAb #76437 (red), Pan-keratin (C11) mouse mAb #4545 (cyan), and LAG3 (D2G4O™) XP® rabbit mAb #15372 (orange).
Immunohistochemistry Image 6: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Multiplex immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using TIM-3 (D5D5R™) XP® rabbit mAb (yellow), PD-1 (D4W2J) XP® rabbit mAb #86163 (green), B7-H3 (D9M2L) XP® rabbit mAb #14058 (red), B7-H4 (D1M8I) XP® rabbit mAb (cyan), LAG3 (D2G4O™) XP® rabbit mAb #15372 (magenta), and VISTA (D1L2G™) XP® rabbit mAb #64953 (orange).
Immunohistochemistry Image 7: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 8

#45208

TIM-3 (D5D5R™) XP® Rabbit mAb

Cell Signaling Technology Logo
Flow cytometric analysis of Jurkat cells (blue) and primary CD4+ T cells (green) using TIM-3 (D5D5R™) XP® Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. CD4+ T cells were purified from human blood and stimulated for 9 days using beads coated with CD3 and CD28 antibodies in the presence of Human Interleukin-2 (hIL-2) #8907 (6.7 ng/ml).
Flow Cytometry Image 1: TIM-3 (D5D5R<sup>™</sup>) XP<sup>®</sup> Rabbit mAb
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.