CD61血小板糖蛋白IIIa（鼠單克隆抗體）

廣州健侖生物科技有限公司

CD61是一個105KDa膜糖蛋白，表達(dá)于血小板、巨核細(xì)胞、破骨細(xì)胞和血管內(nèi)皮細(xì)胞上。CD61可以與整合素β3反應(yīng)，整合素β3鏈接在糖蛋白Ⅱa(CD41)上，形成CD41/CD61復(fù)合物(GPⅡb/Ⅲa)來調(diào)節(jié)血小板的黏附和聚集。CD61主要用于識別血小板及其前驅(qū)細(xì)胞，可用于巨核細(xì)胞白血病的研究。

我司還提供其它進(jìn)口或國產(chǎn)試劑盒：登革熱、瘧疾、流感、A鏈球菌、合胞病毒、腮病毒、乙腦、寨卡、黃熱病、基孔肯雅熱、克錐蟲病、違禁品濫用、肺炎球菌、軍團菌、化妝品檢測、食品安全檢測等試劑盒以及日本生研細(xì)菌分型診斷血清、德國SiFin診斷血清、丹麥SSI診斷血清等產(chǎn)品。

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【產(chǎn)品介紹】

細(xì)胞定位：細(xì)胞漿

克隆號：2f2

同型：IgG1

適用組織：石蠟/冰凍

陽性對照：骨髓巨核細(xì)胞

抗原修復(fù)：熱修復(fù)（EDTA）

抗體孵育時間：30-60min

CD61血小板糖蛋白IIIa（鼠單克隆抗體）

細(xì)胞懸液的制備：取7～8d雄性小鼠5～6只，頸椎脫臼法處死。無菌收集兩側(cè)睪丸，置于盛有PBS的小皿中。整個睪丸收集過程在0.5h內(nèi)完成。用尖鑷子仔細(xì)去除每個睪丸的脂肪墊、附睪及睪丸白膜，加入適量PBS(1～2ml)，吸管用力吹打，將曲細(xì)精管吹散。加入相當(dāng)于組織體積10倍的含1g/L膠原酶的PBS后入溫箱，在37℃、5%CO2條件下作用15min(期間晃動數(shù)次)。之后移入5ml的離心管，輕緩吹打1～2次，靜置待曲細(xì)精管段沉降后輕輕吸除上清，再重復(fù)上述過程1次。加入含1.5g/L透明質(zhì)酸酶和0.25%胰蛋白酶的PBS后入溫箱，同樣條件下作用5～10min，倒置顯微鏡下見曲細(xì)精管段軟散，有的已消散成單細(xì)胞或小的細(xì)胞團即可。加入含10%NBS、1%青、鏈霉素的新鮮D-MEM培養(yǎng)液終止消化，移入離心管中，1 000r/min離心3min或靜置5min，待軟散的曲細(xì)精管及已解離的精原細(xì)胞沉降后，輕輕吸去上清。重新加入1.5ml新鮮培養(yǎng)基(D-MEM+7.5%NBS+7.5%FBS+1%谷氨酰胺+1%非必需氨基酸+1%青、鏈霉素+1%丙酮酸鈉)，吹打8～10次，制成單細(xì)胞懸液。
3.3 Percoll梯度的制備及離心：從已制備好的每級Percoll梯度液中各取1ml，按密度從大到小依次疊加到10ml離心管中。將待分離的細(xì)胞懸液置于梯度zui上層，以1 400r/min離心20min［4］。
3.4 細(xì)胞的洗滌及計數(shù)：用吸管將各梯度中形成的細(xì)胞帶小心取出，移至事先標(biāo)記好的5ml離心管中，各加入1.5ml PBS稀釋Percoll，以1 000r/min的速度離心3min,棄上清，重新加入1.5ml新鮮培養(yǎng)液吹起。取出1滴制備好的細(xì)胞懸液，加入10μl 0.4%臺盼藍(lán)混勻，用紅細(xì)胞計數(shù)板計算活細(xì)胞、死細(xì)胞、細(xì)胞團的數(shù)目和細(xì)胞總數(shù)(沒有吹散的由2個以上細(xì)胞連在一起的細(xì)胞團只記1次)，調(diào)整細(xì)胞密度至3×105個／ml。

我司還提供其它進(jìn)口或國產(chǎn)試劑盒：登革熱、瘧疾、流感、A鏈球菌、合胞病毒、腮病毒、乙腦、寨卡、黃熱病、基孔肯雅熱、克錐蟲病、違禁品濫用、肺炎球菌、軍團菌、化妝品檢測、食品安全檢測等試劑盒以及日本生研細(xì)菌分型診斷血清、德國SiFin診斷血清、丹麥SSI診斷血清等產(chǎn)品。

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【公司名稱】 廣州健侖生物科技有限公司
【市場部】     歐

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【騰訊  】 
【公司地址】 廣州清華科技園創(chuàng)新基地番禺石樓鎮(zhèn)創(chuàng)啟路63號二期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.