BV421 Rat Anti-Human CD366 (TIM-3)

Try the new Anti-Human CD366 antibody: [565562]

BD OptiBuild™ BV421 Rat Anti-Human CD366 (TIM-3)

Name: BV421 Rat Anti-Human CD366 (TIM-3)
Brand: BD OptiBuild™
SKU: 747962

Clone 344823

(RUO)

Flow cytometric analysis of CD366 (TIM-3) expression on peripheral blood leucocytes. Whole blood was treated with 1× BD Pharm Lyse™ Lysing Solution (Cat. No. 555899) to remove erythrocytes. The remaining leucocytes were washed and then costained with PE Mouse Anti-Human CD56 (Cat. No. 555516 or 561903) and either BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602; Left Plots) or BD OptiBuild™ BV421 Rat Anti-Human CD366 (TIM-3) (Cat. No. 747962; Right Plots) at 0.5 μg/test. Left Plots: The pseudocolor dot plots showing the coexpression of CD366 (TIM-3) [or Ig Isotype control staining] versus side-scattered light signals were derived from gated events with the forward and side light scattering characteristics of viable leucocyte populations. Right Plots: The pseudocolor dot plots showing the coexpression of CD366 (TIM-3) [or Ig Isotype control staining] versus CD56 were derived from gated events with the forward and side light scattering characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. The above is qualification data only and does not represent a specific OptiBuild™ lot.

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Product Details

Brand: BD OptiBuild™

Alternative Name: CD366; HAVCR2; T cell immunoglobulin mucin 3; TIM3; TIMD-3; TIMD3; KIM-3

Reactivity: Human (Tested in Development)

Isotype: Rat IgG2a, κ

Immunogen: Human TIM-3 (Ser22-Arg200) Recombinant Protein

Application: Flow cytometry (Qualified)

Concentration: 0.2 mg/ml

Workshop Number: X 10-24

RRID: AB_2872423

Storage Buffer: Aqueous buffered solution containing ≤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 monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.

Recommended Assay Procedures

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.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
Researchers should determine the optimal concentration of this reagent for their individual applications.
An isotype control should be used at the same concentration as the antibody of interest.
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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).

Companion Products

Human BD Fc Block™ RUO

Size 50 µg Cat No. 564219

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Brilliant Stain Buffer RUO

Size 100 Tests Cat No. 563794

Lysing Buffer RUO

Size 100 mL Cat No. 555899

Lysing Solution 10X Concentrate CE_IVD

Size 100 mL Cat No. 349202

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747962 Rev. 2

Antibody Details

344823

The 344823 monoclonal antibody specifically recognizes CD366, which is also known as T cell immunoglobulin mucin 3 (TIM-3), or T-cell immunoglobulin and mucin domain-containing protein 3 (TIMD-3/TIMD3). CD366 (TIM-3) is a ~60 kDa type I transmembrane glycoprotein that is encoded by HAVCR2 (Hepatitis A virus cellular receptor 2) and belongs to the human TIM family (along with TIM-1 and TIM-4) within the Ig superfamily. CD366 (TIM-3) has one Ig-like V-type domain and a mucin stalk region which is rich in serine and threonine. It is expressed on type-1 (Th1-like) CD4+ T cells and CD8+ T cells, type-17 (Th17-like) CD4+ T cells, regulatory T cells (Treg), NKT, and NK cells. It is also expressed on dendritic cells, mast cells, monocytes, macrophages, and microglia but not on B cells or type-2 (Th2-like) CD4+ T cells. CD366 (TIM-3) can regulate macrophage activation and suppress type-1 and type-17 T cell functions. Crosslinking of cell surface CD366 (TIM-3) by binding to Galectin-9 and/or phosphatidylserine can either positively or negatively regulate leucocyte functions, such as cytokine production or the phagocytosis of apoptotic cells. Through its suppressive activity, CD366 (TIM-3) helps maintain peripheral immune tolerance and homeostasis, but can also inhibit anti-tumor immunity.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.

747962 Rev. 2

Format Details

BV421

The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BV421

Excitation Source: Violet 405 nm

Excitation Max: 407 nm

Emission Max: 423 nm

747962 Rev.2

Citations & References

Development References (4)

Cherkassky L, Morello A, Villena-Vargas J, et al. Human CAR T cells with cell-intrinsic PD-1 checkpoint blockade resist tumor-mediated inhibition.. J Clin Invest. 2016; 126(8):3130-44. (Clone-specific: Flow cytometry). View Reference
Monney L1, Sabatos CA, Gaglia JL, et al. Th1-specific cell surface protein Tim-3 regulates macrophage activation and severity of an autoimmune disease.. Nature. 2002; 415(6871):536-541. (Biology). View Reference
Mujib S, Jones RB, Lo C, et al. Antigen-independent induction of Tim-3 expression on human T cells by the common γ-chain cytokines IL-2, IL-7, IL-15, and IL-21 is associated with proliferation and is dependent on the phosphoinositide 3-kinase pathway.. J Immunol. 2012; 188(8):3745-56. (Clone-specific: Flow cytometry). View Reference
Ndhlovu LC, Lopez-Verges S, Barbour JD, et al. Tim-3 marks human natural killer cell maturation and suppresses cell-mediated cytotoxicity. Blood. 2012; 119(16):3734-3743. (Clone-specific: Functional assay). View Reference

View All (4)

747962 Rev. 2

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.