CD61血小板糖蛋白IIIa（鼠单克隆抗体）

广州健仑生物科技有限公司

CD61是一个105KDa膜糖蛋白，表达于血小板、巨核细胞、破骨细胞和血管内皮细胞上。CD61可以与整合素β3反应，整合素β3链接在糖蛋白Ⅱa(CD41)上，形成CD41/CD61复合物(GPⅡb/Ⅲa)来调节血小板的黏附和聚集。CD61主要用于识别血小板及其前驱细胞，可用于巨核细胞白血病的研究。

我司还提供其它进口或国产试剂盒：登革热、疟疾、流感、A链球菌、合胞病毒、腮病毒、乙脑、寨卡、黄热病、基孔肯雅热、克锥虫病、违禁品滥用、肺炎球菌、军团菌、化妆品检测、食品安全检测等试剂盒以及日本生研细菌分型诊断血清、德国SiFin诊断血清、丹麦SSI诊断血清等产品。

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【产品介绍】

细胞定位：细胞浆

克隆号：2f2

同型：IgG1

适用组织：石蜡/冰冻

阳性对照：骨髓巨核细胞

抗原修复：热修复（EDTA）

抗体孵育时间：30-60min

CD61血小板糖蛋白IIIa（鼠单克隆抗体）

细胞悬液的制备：取7～8d雄性小鼠5～6只，颈椎脱臼法处死。无菌收集两侧睾丸，置于盛有PBS的小皿中。整个睾丸收集过程在0.5h内完成。用尖镊子仔细去除每个睾丸的脂肪垫、附睾及睾丸白膜，加入适量PBS(1～2ml)，吸管用力吹打，将曲细精管吹散。加入相当于组织体积10倍的含1g/L胶原酶的PBS后入温箱，在37℃、5%CO2条件下作用15min(期间晃动数次)。之后移入5ml的离心管，轻缓吹打1～2次，静置待曲细精管段沉降后轻轻吸除上清，再重复上述过程1次。加入含1.5g/L透明质酸酶和0.25%胰蛋白酶的PBS后入温箱，同样条件下作用5～10min，倒置显微镜下见曲细精管段软散，有的已消散成单细胞或小的细胞团即可。加入含10%NBS、1%青、链霉素的新鲜D-MEM培养液终止消化，移入离心管中，1 000r/min离心3min或静置5min，待软散的曲细精管及已解离的精原细胞沉降后，轻轻吸去上清。重新加入1.5ml新鲜培养基(D-MEM+7.5%NBS+7.5%FBS+1%谷氨酰胺+1%非必需氨基酸+1%青、链霉素+1%丙酮酸钠)，吹打8～10次，制成单细胞悬液。
3.3 Percoll梯度的制备及离心：从已制备好的每级Percoll梯度液中各取1ml，按密度从大到小依次叠加到10ml离心管中。将待分离的细胞悬液置于梯度zui上层，以1 400r/min离心20min［4］。
3.4 细胞的洗涤及计数：用吸管将各梯度中形成的细胞带小心取出，移至事先标记好的5ml离心管中，各加入1.5ml PBS稀释Percoll，以1 000r/min的速度离心3min,弃上清，重新加入1.5ml新鲜培养液吹起。取出1滴制备好的细胞悬液，加入10μl 0.4%台盼蓝混匀，用红细胞计数板计算活细胞、死细胞、细胞团的数目和细胞总数(没有吹散的由2个以上细胞连在一起的细胞团只记1次)，调整细胞密度至3×105个／ml。

我司还提供其它进口或国产试剂盒：登革热、疟疾、流感、A链球菌、合胞病毒、腮病毒、乙脑、寨卡、黄热病、基孔肯雅热、克锥虫病、违禁品滥用、肺炎球菌、军团菌、化妆品检测、食品安全检测等试剂盒以及日本生研细菌分型诊断血清、德国SiFin诊断血清、丹麦SSI诊断血清等产品。

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【公司名称】 广州健仑生物科技有限公司
【市场部】     欧

【】 
【腾讯  】 
【公司地址】 广州清华科技园创新基地番禺石楼镇创启路63号二期2幢101-103室

Cell suspension preparation: take 7 ~ 8d male mice 5 ~ 6, cervical dislocation method was executed. Both sides of the testes were aseptically collected and placed in small dishes containing PBS. The entire testis collection process completed within 0.5h. Carefully remove the fat pad, epididymis and testis white membrane of each testicle with sharp tweezers, add proper amount of PBS (1 ~ 2ml), force the pipette to blow, and disperse the seminiferous tubules. PBS containing 1 g / L collagenase equivalent to 10 times the volume of the tissue was added to the incubator and allowed to act at 37 ° C under 5% CO 2 for 15 min (shaking several times). After 5ml into the centrifuge tube, gently blow 1 or 2 times, to be fine to be fine tube Segment sedimentation gently sucked the supernatant, and then repeat the above process. Add PBS containing 1.5g / L hyaluronidase and 0.25% trypsin into the incubator under the same conditions for 5 ~ 10min, under inverted microscope, see the seminiferous tubules soft scattered, and some have dissipated into single cells or small The cell mass can be. Add fresh D-MEM broth containing 10% NBS, 1% cyanine and streptomycin to stop digestion, transfer to centrifuge tube, centrifuge at 1000r / min for 3min or stand for 5min, wait for soft scattered seminiferous tubules and solution After the spermatogonia settled, gently aspirate the supernatant. Re-add 1.5ml of fresh medium (D-MEM + 7.5% NBS + 7.5% FBS + 1% Glutamine + 1% nonessential amino acids + 1% cyan, streptomycin + 1% sodium pyruvate) 10 times, made of single cell suspension.
3.3 Preparation of Percoll Gradient and Centrifugation: Take 1ml each from prepared Percoll gradient solution, and add them to 10ml centrifuge tube in descending order of density. The cell suspension to be separated was placed on the top of the gradient and centrifuged at 1400 rpm for 20 min [4].
3.4 cell washing and counting: The gradient formed in the cell straps with a pipette carefully removed, moved to a good mark 5ml centrifuge tube, each adding 1.5ml PBS diluted Percoll, centrifuged at 1 000r / min speed 3min, abandoned Supernatant, re-add 1.5ml fresh medium blown. Remove one drop of the prepared cell suspension, add 10 μl of 0.4% trypan blue and mix. Calculate the number of viable cells, dead cells, cell clusters and total number of cells with the red blood cell counting plate (no more than 2 cells With only one cell group recorded in mind), adjust cell density to 3 × 105 / ml.