CD79a(B细胞)鼠单克隆抗体

广州健仑生物科技有限公司

CD79复合物是异二聚体分子，分别命名Ig-а(CD79a)和Ig-β(CD79b)，与存在于B细胞表面的膜表面免疫球蛋白构成B细胞的抗原识别受体（BCR），参与B细胞活化的信号传导，是一种敏感而特异的B细胞标记物。在B细胞淋巴瘤、大部分的急性B淋巴细胞白血病和骨髓瘤中阳性表达。正常的浆细胞阴性，约50％的浆细胞瘤病例阳性。CD79a作为CD20的一个参考依据，广泛用于B细胞及其来源肿瘤的研究。

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/细胞膜

克隆号：LN2

同型：IgG1

适用组织：石蜡/冰冻

阳性对照：扁桃体

抗原修复：热修复（EDTA）

抗体孵育时间：30-60min

CD79a(B细胞)鼠单克隆抗体

无菌取脾，置于盛有适量无菌Hank''s液的小平皿中，用镊子轻轻将脾磨碎，制成单细胞悬液。经200目筛网过滤，或用4层纱布将脾磨碎，或用Hank''s液洗2次，每次离心10min（1000r/min）。弃上清将细胞浆弹起，加入0.5mL灭菌水20秒，裂解红细胞后再加入0.5mL2倍Hank’s液及8mLHank’s液，1000rpm，10min离心，用1mL含10%小牛血清的RPMI1640*培养液重悬，用1%冰醋酸稀释后计数（活细胞数应在95%以上），用台酚兰染色计数活细胞数（应在95%以上），zui后用RPMI1640*培养液调整细胞浓度为2×107个/mL。
4、NK细胞活性检测
取靶细胞和效应细胞各100μL（效靶比50：1），加入U型96孔培养板中；靶细胞自然释放孔加靶细胞和培养液各100μL，靶细胞zui大释放孔加靶细胞和1％NP40或2.5%Triton各100μL；上述各项均设三个复孔，于37℃、5%CO2培养箱中培养4h，然后将96孔培养板以1500r/min离心5min，每孔吸取上清100μL置平底96孔培养板中，同时加入LDH基质液100μL，反应3min，每孔加入1mol/L的HCl30μL，在酶标仪490nm处测定光密度值（OD）。
按下式计算NK细胞活性，受试样品组的NK细胞活性显著高于对照组的NK细胞活性，即可判定该项实验结果阳性。
                                     反应孔OD－自然释放孔OD
NK细胞活性（%）＝ ────────────────── ×100%
                                   zui大释放孔OD－自然释放孔OD
四、数据处理及结果判定
NK细胞活性需进行数据转换，X＝Sin-1 ，式中P为NK细胞活性，用小数表示，然后再进行方差分析，在进行方差分析时，需按方差分析的程序先进行方差齐性检验，方差齐，计算F值，F值< F0.05，结论：各组均数间差异无显著性；F值≥F0.05，P≤0.05，用多个实验组和一个对照组间均数的两两比较方法进行统计；对非正态或方差不齐的数据进行适当的变量转换，待满足正态或方差齐要求后，用转换后的数据进行统计；若变量转换后仍未达到正态或方差齐的目的，改用秩和检验进行统计。
五、注意事项
1、靶细胞和效应细胞必须新鲜，细胞存活率应大于95％。
2、比色时环境温度应保持恒定。
3、LDH基质液应临用前配制。
4、在一定范围内，NK细胞活性与效靶比值成正比。一般效靶比值不应超过100。

CD79a(B细胞)鼠单克隆抗体

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。

想了解更多的产品及服务请扫描下方二维码：

【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

Aseptically take the spleen, placed in a small amount of sterile Hank's' s dish, gently tweezers tweezers to make a single cell suspension. Filter through a 200-mesh sieve, or grind the spleen with 4 layers of gauze or wash twice in Hank's's for 10 min each (1000 r / min). The supernatant was discarded supernatant of cytoplasm, adding 0.5mL sterile water for 20 seconds, red blood cells were lysed after adding 0.5mL2 times Hank's solution and 8mL Hank's solution, 1000rpm, 10min centrifugation, with 1mL of 10% fetal calf serum RPMI1640 complete medium Resuspended, diluted with 1% glacial acetic acid (live cell number should be above 95%), count the number of viable cells (should be above 95%) with Phenol blue staining, and finally adjust the cell concentration to 2 with RPMI1640 complete medium × 107 / mL.
4, NK cell activity test
Taking target cells and effector cells in each 100 L (effector to target ratio of 50: 1), was added a U-shaped 96-well culture plate; target cell spontaneous release well of target cells and culture of each 100 L solution, maximum release holes target cells plus target cells and 1 % NP40 or 100 L each of 2.5% Triton; the above three wells are located at 37 ℃, 5% CO2 incubator for 4h, then the 96-well culture plates at 1500r / min centrifugal 5min, supernatant of each well to draw 100μL flat-bottom 96-well culture plate, while adding 100μL LDH matrix solution, the reaction 3min, 1mol / L HCl was added to each well 30μL, measured at a microplate reader 490nm optical density (OD).
NK cell activity was calculated according to the following formula, NK cell activity in the test sample group was significantly higher than that in the control group NK cell activity, you can determine the positive results of the experiment.
                                     Reaction Hole OD - Natural Release Hole OD
NK cell activity (%) = ────────────────── × 100%
                                   Maximum release hole OD - Natural release hole OD
Fourth, data processing and result judgment
NK cell activity program required for data conversion, X = Sin-1, where P is the NK cell activity, as a decimal number, then the analysis of variance, performed the analysis of variance, analysis of variance basis having to test for homogeneity of variance , The variance of Qi, F value calculation, F value <F0.05, Conclusion: There was no significant difference between the mean of each group; F value ≥ F0.05, P≤0.05, using a number of experimental groups and a control group pairwise statistical comparison method; non-normality of the data or the heterogeneity of variance appropriate variable conversion, to be satisfied after a normal or variance homogeneity requirements, the statistical data conversion; after conversion if the variable n has not yet reached The purpose of the state or variance Qi, using rank sum test statistics.
Five, matters needing attention
1, target cells and effector cells must be fresh, cell survival rate should be greater than 95%.
2, the color temperature should be kept constant.
3, LDH matrix solution should be prepared before use.
4, within a certain range, NK cell activity is proportional to the ratio of effective target. General target ratio should not exceed 100.