CD79a(B細(xì)胞)鼠單克隆抗體

廣州健侖生物科技有限公司

CD79復(fù)合物是異二聚體分子,，分別命名Ig-а(CD79a)和Ig-β(CD79b),，與存在于B細(xì)胞表面的膜表面免疫球蛋白構(gòu)成B細(xì)胞的抗原識別受體（BCR）,，參與B細(xì)胞活化的信號傳導(dǎo),，是一種敏感而特異的B細(xì)胞標(biāo)記物,。在B細(xì)胞淋巴瘤、大部分的急性B淋巴細(xì)胞白血病和骨髓瘤中陽性表達(dá),。正常的漿細(xì)胞陰性,，約50％的漿細(xì)胞瘤病例陽性。CD79a作為CD20的一個參考依據(jù),，廣泛用于B細(xì)胞及其來源腫瘤的研究,。

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【產(chǎn)品介紹】

細(xì)胞定位：細(xì)胞漿/細(xì)胞膜

克隆號：LN2

同型：IgG1

適用組織：石蠟/冰凍

陽性對照：扁桃體

抗原修復(fù)：熱修復(fù)（EDTA）

抗體孵育時間：30-60min

CD79a(B細(xì)胞)鼠單克隆抗體

無菌取脾，置于盛有適量無菌Hank''s液的小平皿中,，用鑷子輕輕將脾磨碎,，制成單細(xì)胞懸液。經(jīng)200目篩網(wǎng)過濾,，或用4層紗布將脾磨碎,，或用Hank''s液洗2次，每次離心10min（1000r/min）,。棄上清將細(xì)胞漿彈起,，加入0.5mL滅菌水20秒，裂解紅細(xì)胞后再加入0.5mL2倍Hank’s液及8mLHank’s液,，1000rpm,，10min離心，用1mL含10%小牛血清的RPMI1640*培養(yǎng)液重懸,，用1%冰醋酸稀釋后計數(shù)（活細(xì)胞數(shù)應(yīng)在95%以上）,，用臺酚蘭染色計數(shù)活細(xì)胞數(shù)（應(yīng)在95%以上），zui后用RPMI1640*培養(yǎng)液調(diào)整細(xì)胞濃度為2×107個/mL,。
4,、NK細(xì)胞活性檢測
取靶細(xì)胞和效應(yīng)細(xì)胞各100μL（效靶比50：1），加入U型96孔培養(yǎng)板中,；靶細(xì)胞自然釋放孔加靶細(xì)胞和培養(yǎng)液各100μL,，靶細(xì)胞zui大釋放孔加靶細(xì)胞和1％NP40或2.5%Triton各100μL；上述各項均設(shè)三個復(fù)孔,，于37℃,、5%CO2培養(yǎng)箱中培養(yǎng)4h，然后將96孔培養(yǎng)板以1500r/min離心5min,，每孔吸取上清100μL置平底96孔培養(yǎng)板中,，同時加入LDH基質(zhì)液100μL，反應(yīng)3min,，每孔加入1mol/L的HCl30μL,，在酶標(biāo)儀490nm處測定光密度值（OD）。
按下式計算NK細(xì)胞活性,，受試樣品組的NK細(xì)胞活性顯著高于對照組的NK細(xì)胞活性,，即可判定該項實驗結(jié)果陽性。
                                     反應(yīng)孔OD－自然釋放孔OD
NK細(xì)胞活性（%）＝ ────────────────── ×100%
                                   zui大釋放孔OD－自然釋放孔OD
四,、數(shù)據(jù)處理及結(jié)果判定
NK細(xì)胞活性需進(jìn)行數(shù)據(jù)轉(zhuǎn)換,，X＝Sin-1 ，式中P為NK細(xì)胞活性,，用小數(shù)表示,，然后再進(jìn)行方差分析,，在進(jìn)行方差分析時，需按方差分析的程序先進(jìn)行方差齊性檢驗,，方差齊,，計算F值，F(xiàn)值< F0.05,，結(jié)論：各組均數(shù)間差異無顯著性,；F值≥F0.05，P≤0.05,，用多個實驗組和一個對照組間均數(shù)的兩兩比較方法進(jìn)行統(tǒng)計,；對非正態(tài)或方差不齊的數(shù)據(jù)進(jìn)行適當(dāng)?shù)淖兞哭D(zhuǎn)換，待滿足正態(tài)或方差齊要求后,，用轉(zhuǎn)換后的數(shù)據(jù)進(jìn)行統(tǒng)計,；若變量轉(zhuǎn)換后仍未達(dá)到正態(tài)或方差齊的目的，改用秩和檢驗進(jìn)行統(tǒng)計,。
五,、注意事項
1、靶細(xì)胞和效應(yīng)細(xì)胞必須新鮮,，細(xì)胞存活率應(yīng)大于95％,。
2、比色時環(huán)境溫度應(yīng)保持恒定,。
3,、LDH基質(zhì)液應(yīng)臨用前配制。
4,、在一定范圍內(nèi)，NK細(xì)胞活性與效靶比值成正比,。一般效靶比值不應(yīng)超過100,。

CD79a(B細(xì)胞)鼠單克隆抗體

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(tuán)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

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.