Biolegend, 285020, Spark Blue™ 574 anti-mouse CD90.2 (Thy1.2) (Flexi-Fluor™) Antibody, 50μg

CD90.2 is a 25-35 kD immunoglobulin superfamily member also known as Thy1.2. It is expressed on hematopoietic stem cells and neurons, all thymocytes, and peripheral T cells in Thy1.2 bearing mouse strains (Balb/c, CBA/J, C3H/He, C57BL/-, DBA, NZB/-). CD90.2 is a glycosylphosphatidylinositol (GPI)-anchored membrane glycoprotein involved in signal transduction. CD90.2 is involved in costimulation of lymphocyte proliferation and induction of hematopoietic stem cells differentiation. CD90.2 has been shown to interact with CD45. The 30-H12 antibody has been reported to induce Ca 2+ flux in thymocytes and, in combination with antibody against the CD3/TCR complex, promote thymocyte apoptosis and inhibit CD3-mediated proliferative responses of mature T lymphocytes.
50μg
Verified Reactivity: Mouse
Antibody Type: Monoclonal
Host Species: Rat
Immunogen: Mouse thymus or spleen
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation: The antibody was purified by affinity chromatography and conjugated with Spark Blue™ 574 under optimal conditions.
Concentration: 0.2 mg/mL
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application: FC
Recommended Usage: Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL. For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration. * Spark Blue™ 574 has a maximum excitation of 506 nm and a maximum emission of 574 nm.
Excitation Laser: Blue Laser (488 nm)
Application Notes: Additional reported applications (for the relevant formats) include: in vivo and in vitro depletion1,2,7, costimulation of CD3/TCR-mediated signal transduction3,4, and immunohistochemical staining5 of acetone-fixed frozen sections. The 30-H12 antibody does not react with Thy-1.1 alloantigen of the AKR/J and PL strains. To reduce non-specific binding to cells bearing Fc-receptors, pre-incubation of cells with anti-mouse CD16/CD32, clone 93 (Cat. No. 101301 & 101302) is recommended prior to immunofluorescent staining. The Ultra-LEAF™ purified antibody (Endotoxin <0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. Nos. 105351 & 105352).
Additional Product Notes: For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.
Application References(PubMed link indicates BioLegend citation): Hathcock KS. 1991. Current Protocols in Immunology. 3.4.1. (Deplete) Seaman WE. 1983. J. Immunol. 130:1713. (Deplete) Nakashima I, et al. 1991. J. Immunol. 147:1153. (Costim) Nakashima I, et al. 1993. J. Immunol. 151:3511. (Costim) Ledbetter JA, et al. 1980. J. Exp. Med. 152:280. (IHC) Hardy B, et al. 2005. Int. Immunol. 17:615. Drobyski W, et al. 1996. Blood 87:5355. (Deplete) Dyer KD, et al. 2007. J. Immunol. 179:1693. (FC) PubMed Sungur CM, et al. 2013. PNAS. 110:7401. PubMed
RRID: AB_3106285 (BioLegend Cat. No. 285020)
Structure: Ig superfamily, 25-35 kD
Distribution: Hematopoietic stem cells and neurons, all thymocytes, peripheral T cells of the Thy-1.2 bearing mice
Function: Lymphocyte costimulation, proliferation and differentiation of hematopoietic stem cells
Ligand/Receptor: CD45
Cell Type: Hematopoietic stem and progenitors, Neurons, T cells, Thymocytes
Biology Area: Immunology
Molecular Family: CD Molecules
Antigen References: 1. Barclay A, et al. 1997. The Leukocyte Antigen FactsBook Academic Press. 2. Craig W, et al. 1993. J. Exp. Med. 177:1331. 3. Reif AE and Schlesinger M. 1989. Cell Surface Antigen Thy-1. 4. Mayani H, et al. 1994. Blood 83:2410.
Gene ID: 21838
UniProt: View information about CD90.2 on UniProt.org
Clone: 30-H12
Regulatory Status: RUO
Other Names: Thy-1.2
Isotype: Rat IgG2b, κ
Q: What are Flexi-Fluors?
A: Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer.
Q: How quickly will I receive my order?
A: We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary.
Q: How are Flexi-Fluors different from regular catalog products?
A: Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.
Q: How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?
A: Flexi-Fluors are provided at a standard 0.2 mg/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg/mL. 

For example, make five 1:1 serial dilutions of your 0.2 mg/mL antibody.  Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.
Q: I can’t find the antibody-dye combination that I need. When will it be available?
A: We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services.
Q: I need help to validate the performance of my Flexi-Fluor. Who should I contact?
A: Please get in touch with Technical Service for assistance.
Q: Can I order more than 50 μg of a Flexi-Fluor?
A: Yes, you can order multiple vials of the same Flexi-Fluor products.  We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.
Q: What is the expiration date of my Flexi-Fluor?
A: Expiration dates can be found on the vial label or by using our CoA lookup tool.
Q: What if my requested antibody-dye combination doesn't perform as expected?
A: Flexi-Fluor™ products have been carefully selected by our scientists for their excellent conjugation properties and broad compatibility. In the rare case that an antibody-dye combination does not perform as expected due to the unique characteristics of a specific pairing, please contact our Technical Service team. We will work with you to identify alternative options to meet your panel requirements.