PE-CF594 Mouse Anti-Mouse CD366 (TIM-3)

BD Horizon™ PE-CF594 Mouse Anti-Mouse CD366 (TIM-3)

Name: PE-CF594 Mouse Anti-Mouse CD366 (TIM-3)
Brand: BD Horizon™
SKU: 566998

クローン 5D12/TIM-3 (also known as 5D12)

(RUO)

Flow cytometric analysis of CD366 (TIM-3) expressed on activated mouse splenocytes. BALB/c splenic leucocytes were cultured for 4 days in the presence of plate-bound Purified NA/LE Hamster Anti-Mouse CD3e (Cat. No. 553057; 10 μg/ml for coating), soluble Purified NA/LE Hamster Anti-Mouse CD28 (Cat. No. 553294; 2 μg/ml), and Purified NA/LE Rat Anti-Mouse IL-4 (Cat. No. 554432; 1 μg/ml) antibodies. The cells were harvested, preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142), and then stained with BD Horizon™ BV421 Rat Anti-Mouse CD8a antibody (Cat. No. 563898) and either BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; Left Plot) or BD Horizon PE-CF594 Mouse Anti-Mouse CD366 (TIM-3) antibody (Cat. No. 566998; Right Plot) at 0.5 μg/test. BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) was added to cells right before analysis. A bivariate pseudocolor density plot showing the correlated expression of CD8a versus CD366 (TIM-3) [or Ig Isotype control staining] was derived from gated events with the forward and side light-scatter characteristics of viable (7-AAD-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Spectrum Viewer

Protocol Library

Scientific Resources

製品詳細

Brand: BD Horizon™

Alternative Name: Cd366; T-cell membrane protein; Tim3; TIMD-3; Timd3; HAVcr-2; Havcr2;

Reactivity: Mouse (QC Testing)

Isotype: Mouse IgG1, κ

Immunogen: Mouse TIM-3 Recombinant Protein

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

RRID: AB_2869998

Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation and Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated with BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

推奨アッセイ手順

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
CF™ is a trademark of Biotium, Inc.
This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
An isotype control should be used at the same concentration as the antibody of interest.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Since applications vary, each investigator should titrate the reagent to obtain optimal results.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.

サイズ 0.1 mg カタログ番号 553141

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO

サイズ 0.5 mg カタログ番号 553142

Stain Buffer (FBS) RUO

サイズ 500 mL カタログ番号 554656

Stain Buffer (BSA) RUO

サイズ 500 mL カタログ番号 554657

Lysing Buffer RUO

サイズ 100 mL カタログ番号 555899

PE-CF594 Mouse IgG1, κ Isotype Control RUO

サイズ 0.1 mg カタログ番号 562292

詳細を表示

566998 Rev. 1

抗体の詳細

5D12/TIM-3

The 5D12 monoclonal antibody specifically recognizes CD366 which is also known as TIM-3 (T-cell immunoglobulin and mucin domain-containing 3) or T-cell membrane protein 3. TIM-3 is encoded by Havcr2 (hepatitis A virus cellular receptor 2). TIM-3 is a type I transmembrane glycoprotein that belongs to the human TIM family within the immunoglobulin superfamily, having one Ig-like V-type domain in its extracellular region. TIM-3 is expressed on activated monocytes, macrophages, dendritic cells, microglia, and mast cells. It is also expressed on type-1 CD4+ (Th1-like) T cells, CD8+ T cytotoxic cells, regulatory T cells (Treg), and natural killer (NK) cells. TIM-3 functions as an inhibitory receptor which helps cells maintain immunological homeostasis and self tolerance. Crosslinking of cell surface TIM-3 by Galectin-9 binding downregulates Th1-like and CD8+ T cell responses and can promote Treg or myeloid-derived suppressor cells. TIM-3 enables dendritic cells to bind phosphatidyl serine expressed by apoptotic cells and to phagocytize these cells to quell potential inflammation. TIM-3 also binds to the alarmin HMGB1 thereby preventing Toll-like receptor (TLR) by released tumor cell DNA.

This antibody is conjugated to BD Horizon PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg, 610/20-nm filter).

566998 Rev. 1

フォーマットの詳細

PE-CF594

BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

フォーマット: PE-CF594

励起源: Yellow-Green 488 nm, 532 nm, 561 nm

最大励起波長: 496 nm, 566 nm

最大蛍光波長: 615 nm

566998 Rev.1

引用＆参考文献

Development References (4)

Anderson AC, Joller N, Kuchroo VK. Lag-3, Tim-3, and TIGIT: Co-inhibitory Receptors with Specialized Functions in Immune Regulation.. Immunity. 2016; 44(5):989-1004. (Biology). View Reference
Oikawa T1, Kamimura Y, Akiba H, et al. Preferential involvement of Tim-3 in the regulation of hepatic CD8+ T cells in murine acute graft-versus-host disease.. J Immunol. 2006; 177(7):4281-4287. (Biology). View Reference
Phong BL, Avery L, Sumpter TL, et al. Tim-3 enhances FcεRI-proximal signaling to modulate mast cell activation.. J Exp Med. 2015; 212(13):2289-304. (Clone-specific: Flow cytometry). View Reference
Veenstra RG, Taylor PA, Zhou Q, et al. Contrasting acute graft-versus-host disease effects of Tim-3/galectin-9 pathway blockade dependent upon the presence of donor regulatory T cells.. Blood. 2012; 120(3):682-90. (Clone-specific: Flow cytometry). View Reference

すべて表示する (4)

566998 Rev. 1

Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

BD Horizon™ PE-CF594 Mouse Anti-Mouse CD366 (TIM-3)

Preparation and Storage

推奨アッセイ手順

Product Notices

関連製品

Development References (4)