{"product_id":"t-cell-activation-nanobeadshumanruo-s0k0020","title":"T Cell Activation Nanobeads，Human（RUO）","description":"\u003ch4\u003eProduct Specification\u003c\/h4\u003e\u003cdiv class=\"responsive-table product-detail-table details-table\"\u003e\n\u003cbr\u003e\u003ctable style=\"width: 100%; height: auto;\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eFormat\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\n\u003cp\u003e\u003cspan\u003eDextran-coated iron tetroxide nanobeads with surface-coupled anti-human CD3 and anti-human CD28 specific antibodies;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e1 mL Human T Cell Activation Nanobeads: can activate 2 × 10⁸ T cells;\u003cbr\u003e0.5 mL Human T Cell Activation Nanobeads: can activate 1 × 10⁸ T cells.\u003cbr\u003e\u003c\/p\u003e\n\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eReactivity\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003eHuman\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eMagnetic Bead Size\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e20 nm\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eEndotoxin\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u0026lt;1EU\/mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 22%;\"\u003e\u003cstrong\u003eStability \u0026amp; Storage\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"width: 78%;\"\u003e\u003cp\u003eStore protected from light at 2–8 °C; do not freeze.\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003c\/div\u003e\u003ch4\u003eBackground\u003c\/h4\u003e\u003cdiv\u003e\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"color:#000000\"\u003e \u003c\/span\u003e\u003c\/p\u003e\u003c\/div\u003e\u003ch4\u003eProtocol\u003c\/h4\u003e\u003cdiv\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1.\u003c\/span\u003e\u003cspan style=\"font-weight:bold;color:#000000\"\u003ePrecautions\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eAll operations throughout the entire process must be performed in a sterile environment;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eFor initial activation, ensure that the reagent continuously interacts with the cells for 2–3 days; removing the reagent too early will result in insufficient T-cell activation and reduced proliferative capacity;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eT cells can undergo viral transduction experiments two days after activation; the optimal transduction titer and timing should be determined independently through experimentation;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eResidual EDTA in the cell system significantly inhibits T-cell activation stimulation; it must be thoroughly removed before starting the experiment.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold;color:#000000\"\u003e2. Sample Preparation\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"color:#000000\"\u003eCD4 Nanobeads, human (Cat. No. S0K0013), CD8 Nanobeads, human (Cat. No. S0K0005), and CD3 Nanobeads, human (Cat. No. S0K0011) can be used to isolate and enrich T cells from PBMCs. Refer to the corresponding product manuals for specific procedures.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"color:#000000\"\u003ePeripheral blood samples from humans need to be separated by density gradient centrifugation to obtain PBMCs, which are then used for subsequent T-cell activation experiments.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold;color:#000000\"\u003e3. Cell Seeding Density and Reagent Usage Standards\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"color:#000000\"\u003eThis experimental protocol optimally recommends using a working dilution of 1:200; the best seeding density for sorted T cells is 1×10⁶ cells\/cm²; the optimal seeding density for PBMCs is 2×10⁶ cells\/cm².\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"color:#000000\"\u003eOptimal Culture Medium Formulation Table for Each Type of Culture Plate\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" style=\"border-collapse:collapse;width:auto;border:1px solid #000000\"\u003e\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold;color:#000000\"\u003eCulture Plate Model\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold;color:#000000\"\u003eGrowth Surface Area\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold;color:#000000\"\u003eTotal Culture Volume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold;color:#000000\"\u003eT-Cell Inoculum Quantity\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold;color:#000000\"\u003eNanobeads Addition Amount\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"font-weight:bold;color:#000000\"\u003eCell Culture Medium Volume\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e96-well plate\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e0.31 cm²\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e0.2 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e0.3 × 10⁶\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e1 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e199 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e48-well plate\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e1 cm²\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e1 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e1 × 10⁶\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e5 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e995 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e24-well plate\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e2 cm²\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e2 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e2 × 10⁶\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e10 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e1990 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e12-well plate\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e4 cm²\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e4 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e4×10⁶\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e20 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e3980 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e6-well plate\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e10 cm²\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e5 mL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e5×10⁶\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e25 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align:center;vertical-align:middle\"\u003e\u003cp style=\"text-align:center\"\u003e\u003cspan style=\"color:#000000\"\u003e4975 μL\u003c\/span\u003e\u003c\/p\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\u003c\/table\u003e\n\u003cp style=\"margin-bottom: 0px;\"\u003e\u003cbr\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"color:#000000\"\u003eNotes: When the cell count is less than 10⁶, still add reagents in fixed volumes as per the table; when scaling up the cell culture, proportionally increase the total volumes of reagents and culture medium.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold;color:#000000\"\u003e4. Standard activation procedure for a 48-well plate\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eUse AO\/PI to count cells and determine the required cell amount;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eResuspend the cells in 995 μL of complete T-cell culture medium;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eAdd 5 μL\u003c\/span\u003e\u003cspan\u003eT Cell Activation Nanobeads\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eand mix gently and thoroughly;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003ePlace in a 37℃, 5% CO₂ incubator and let it sit for 48 hours;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eAfter the initial activation, perform centrifugation and media exchange:\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"color:#000000\"\u003e① Centrifuge at 300×g for 10 min;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"color:#000000\"\u003e② Slowly and completely aspirate the supernatant from the upper layer;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"color:#000000\"\u003e③ Add fresh T cell complete culture medium at a concentration of 1×10⁶\/ml;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"color:#000000\"\u003e④ Incubate in a 37℃, 5% CO₂ incubator to continue culturing.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"font-weight:bold;color:#000000\"\u003e5. Cell expansion and culture procedure\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e1)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eEvery 2 days, equally divide the cell suspension into new wells or flasks, and replenish with fresh culture medium;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e2)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eMaintain standard culture conditions at 37℃ and 5% CO₂;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e3)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eBy day 12 of culture, the cells have expanded to sufficient numbers and can be used for various downstream experiments;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e4)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eDepending on the cells' growth status and experimental design requirements, the expansion culture period can be extended up to 16 days at most;\u003c\/span\u003e\u003c\/p\u003e\n\u003cp style=\"text-align:left\"\u003e\u003cspan style=\"margin-right: 8px;\"\u003e5)\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003eIf long-term expansion culture is needed, T Cell Activation Nanobeads can be added again after day 12 of culture.\u003c\/span\u003e\u003cspan\u003eT Cell Activation Nanobeads\u003c\/span\u003e\u003cspan style=\"color:#000000\"\u003ePerform secondary activation stimulation.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e","brand":"Starter","offers":[{"title":"0.5 mL","offer_id":43030516236363,"sku":"S0K0020-0.5 mL","price":0.1,"currency_code":"USD","in_stock":true},{"title":"1 mL","offer_id":43030516269131,"sku":"S0K0020-1 mL","price":0.1,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0590\/8375\/1499\/files\/a3b06f5569264f0ea9099f1ed448d80d.jpg?v=1786510869","url":"https:\/\/www.antbioinc.com\/products\/t-cell-activation-nanobeadshumanruo-s0k0020","provider":"AntBio","version":"1.0","type":"link"}