{"product_id":"biolegend-300446","title":"Biolegend, 300446, Alexa Fluor® 594 anti-human CD3 Antibody, 100μg","description":"\u003cp\u003eCD3ε is a 20 kD chain of the CD3\/T-cell receptor (TCR) complex which is composed of two CD3ε, one CD3γ, one CD3δ, one CD3ζ (CD247), and a T-cell receptor (α\/β or γ\/δ) heterodimer. It is found on all mature T cells, NKT cells, and some thymocytes. CD3, also known as T3, is a member of the immunoglobulin superfamily that plays a role in antigen recognition, signal transduction, and T cell activation.\u003cbr\u003e\n100μg\u003cbr\u003e\nVerified Reactivity: Human\u003cbr\u003e\nReported Reactivity: Chimpanzee\u003cbr\u003e\nAntibody Type: Monoclonal\u003cbr\u003e\nHost Species: Mouse\u003cbr\u003e\nFormulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.\u003cbr\u003e\nPreparation: The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 594 under optimal conditions.\u003cbr\u003e\nConcentration: 0.5 mg\/mL\u003cbr\u003e\nStorage \u0026amp; Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.\u003cbr\u003e\nApplication: ICC - Quality tested FC - Verified SB - Reported in the literature, not verified in house\u003cbr\u003e\nRecommended Usage: Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 2.5 - 10 μg\/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application. * Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm. Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.View full statement regarding label licenses\u003cbr\u003e\nExcitation Laser: Green Laser (532 nm)\/Yellow-Green Laser (561 nm)\u003cbr\u003e\nApplication Notes: Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen sections4,6,7 and formalin-fixed paraffin-embedded sections11, immunoprecipitation1, activation of T cells2,3,5, Western blotting9, and spatial biology (IBEX)16,17. The LEAF™ purified antibody (Endotoxin \u0026lt; 0.1 EU\/µg, Azide-Free, 0.2 µm filtered) is recommended for functional assays (Cat. No. 300413, 300414, and 300432). For highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 300437, 300438, 300465, 300466, 300473, 300474) with a lower endotoxin limit than standard LEAF™ purified antibodies (Endotoxin \u0026lt; 0.01 EU\/µg).\u003cbr\u003e\nAdditional Product Notes: Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH). View more applications data for this product in our Scientific Poster Library.\u003cbr\u003e\nApplication References(PubMed link indicates BioLegend citation): Salmeron A, et al. 1991. J. Immunol. 147:3047. (IP) Graves J, et al. 1991. J. Immunol. 146:2102. (Activ) Lafont V, et al. 2000. J. Biol. Chem. 275:19282. (Activ) Ryschich E, et al. 2003. Tissue Antigens 62:48. (IHC) Thompson AG, et al. 2004. J. Immunol. 173:1671. (Activ) Sakkas LI, et al. 1998. Clin. Diagn. Lab. Immun. 5:430. (IHC) Mack CL, et al. 2004. Pediatr. Res. 56:79. (IHC) Thakral D, et al. 2008. J. Immunol. 180:7431. (FC) PubMed Van Dongen JJM, et al. 1988. Blood 71:603. (WB) Yoshino N, et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC) Pollard, K. et al. 1987. J. Histochem. Cytochem. 35:1329. (IHC) Luckashenak N, et al. 2013. J. Immunol. 190:27. PubMed Laurent AJ, et al. 2014. PLoS One. 9:103683. PubMed Li J, et al. 2015. Cancer Res. 75:508. PubMed Stoeckius M, et al. 2017. Nat. Methods. 14:865-868. (PG) Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed\u003cbr\u003e\nProduct Citations: Emmons TR, et al. 2021. Cancer Immunol Res. 9:790. PubMed Mehta AK, et al. 2021. Nat Cancer. 2:66. PubMed Su Y, et al. 2021. Cancers (Basel). 13:. PubMed\u003cbr\u003e\nRRID: AB_2563236 (BioLegend Cat. No. 300446)\u003cbr\u003e\nStructure: Ig superfamily, with the subunits of CD3γ, CD3δ, CD3ζ (CD247) and TCR (α\/β or γ\/δ) forms CD3\/TCR complex, 20 kD\u003cbr\u003e\nDistribution: Mature T and NK T cells, thymocyte differentiation\u003cbr\u003e\nFunction: Antigen recognition, signal transduction, T cell activation\u003cbr\u003e\nLigand\/Receptor: Peptide antigen bound to MHC\u003cbr\u003e\nCell Type: NKT cells, T cells, Thymocytes, Tregs\u003cbr\u003e\nBiology Area: Immunology, Innate Immunity\u003cbr\u003e\nMolecular Family: CD Molecules, TCRs\u003cbr\u003e\nAntigen References: 1. Barclay N, et al. 1993. The Leucocyte FactsBook. Academic Press. San Diego. 2. Beverly P, et al. 1981. Eur. J. Immunol. 11:329. 3. Lanier L, et al. 1986. J. Immunol. 137:2501-2507.\u003cbr\u003e\nGene ID: 916\u003cbr\u003e\nUniProt: View information about CD3 on UniProt.org\u003cbr\u003e\nClone: UCHT1\u003cbr\u003e\nRegulatory Status: RUO\u003cbr\u003e\nWorkshop: III 471\u003cbr\u003e\nOther Names: T3, CD3ε\u003cbr\u003e\nIsotype: Mouse IgG1, κ\u003cbr\u003e\nQ: If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?\u003cbr\u003e\nA: It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.\u003cbr\u003e\nQ: Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?\u003cbr\u003e\nA: Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF\/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.\u003cbr\u003e\nQ: Are other fluorophores compatible with IBEX?\u003cbr\u003e\nA: Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.\u003cbr\u003e\nQ: The same antibody works in one tissue type but not another. What is happening?\u003cbr\u003e\nA: Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.\u003cbr\u003e\nQ: How can I be sure the staining I’m seeing in my tissue is real?\u003cbr\u003e\nA: In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.\u003c\/p\u003e","brand":"Biolegend","offers":[{"title":"Default Title","offer_id":46864074408105,"sku":"300446","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/es\/products\/biolegend-300446","provider":"Iright","version":"1.0","type":"link"}