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甲型乙型流感診斷試劑盒(快檢方法)
廣州健侖生物科技有限公司
廣州健侖長期供應(yīng)各種流感檢測試劑,包括進(jìn)口和國產(chǎn)的品牌,主要包括日本富士瑞必歐、日本生研、美國BD、美國NovaBios、美國binaxNOW、凱必利、廣州創(chuàng)侖等主流品牌。
甲型乙型流感診斷試劑盒(快檢方法)
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【公司名稱】 廣州健侖生物科技有限公司
【市場部】 楊永漢
【】
【騰訊 】 2042552662
【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(hào)二期2幢101-103室
二甲苯
酒精(100%,95%)
EDTA抗原修復(fù)工作液(pH8.0,稀釋50倍使用)
0.5M Tris-NaCl (TBS) pH7.4(稀釋10倍使用)
DAB顯色液(臨用前配制:試劑盒A、B、C各50ul,于1ml水中混勻)。
方法
1. 石蠟切片置60℃烘箱中烘烤過夜
2. 二甲苯中脫蠟,梯度酒精入水(無水乙醇,95%酒精),浸泡于蒸餾水中待用
3. 抗原修復(fù)
取500mlEDTA抗原修復(fù)工作液于1000ml燒杯中,在小功率電爐上加熱,至似沸微沸(為了防止脫片)。將組織切片緩慢放入燒杯。繼續(xù)加熱,保持液體在微沸狀態(tài)20分鐘。將燒杯移開火源,室溫下自然冷卻后取出切片,蒸餾水洗1次3分鐘,TBS洗2次每次3分鐘。
4. 每張切片加一滴或50ul 3%過氧化氫溶液,室溫下孵育10min, 以阻斷內(nèi)源性過氧化物酶的活性。 TBS沖洗3次,每次3min。
5. 除去TBS液,加一滴或50ul 一抗(滅活PLB免疫小鼠所得到的多抗,1:3000稀釋),陰性對照采用普通血清, 室溫下孵育2小時(shí),或4℃過夜。
6. TBS洗3次,每次5min,除去TBS液,每張切片加一滴或50ul polymer enhancer(試劑A), 室溫下孵育20min, TBS沖洗3次, 每次3min。
7. 除去TBS液,每張切片加一滴或50ul 過氧化物酶標(biāo)記的抗鼠/兔聚合物(試劑B),室溫下孵育30min,TBS洗3次,每次3min。
8. 除去TBS液,每張切片加一滴或50ul 新鮮配制的DAB,顯色3-10min。
9. 自來水沖洗,蘇木素復(fù)染10min,水沖洗。0.1%的鹽酸分化,自來水沖洗,TBS返藍(lán)。
10. 不需脫水,直接用中性樹脂封片。
Xylene
Alcohol (100%, 95%)
EDTA antigen repair working solution (pH8.0, diluted 50 times use)
0.5M Tris-NaCl (TBS) pH7.4 (diluted 10 times used)
DAB color reagent (before preparation: kit A, B, C of each 50ul, 1ml water mix).
method
1. Paraffin slices were oven-baked overnight at 60 ° C
2. Xylene dewaxing, gradient alcohol into the water (ethanol, 95% alcohol), soaked in distilled water to be used
Antigen repair
Take 500mlEDTA antigen repair working solution in 1000ml beaker, heated in a low-power electric furnace, to the boiling-like micro boiling (in order to prevent peeling). Slowly place tissue sections into beakers. Continue to heat, keep the liquid in a slightly boiling state for 20 minutes. The beaker is removed from the source of fire, cooled at room temperature and then remove the slice, washed with distilled water 3 minutes 3 times, TBS wash 2 times each 3 minutes.
4. Add one drop or 50μl of 3% hydrogen peroxide solution to each slice and incubate at room temperature for 10min to block the activity of endogenous peroxidase. TBS rinse 3 times, each 3min.
5. Remove the TBS solution and add one drop or 50 ul of primary antibody (1: 3000 dilution of polyclonal antibody obtained from mice immunized with PLB). The negative control was incubated with normal serum for 2 hours at room temperature or 4 ° C overnight.
6. TBS wash three times, each time 5min, remove the TBS solution, add one drop or 50ul of polymer enhancer per slice (Reagent A), incubate 20min at room temperature, TBS rinse 3 times, each 3min.
7. Remove the TBS solution by adding one drop or 50ul of peroxidase-labeled anti-mouse / rabbit polymer (Reagent B) to each section. Incubate for 30 min at room temperature and 3x TBS for 3 min each.
8. Remove the TBS solution, add one drop or 50ul of freshly prepared DAB per slice, and color develop 3-10min.
9. Tap water, hematoxylin counterstain 10min, water rinse. 0.1% hydrochloric acid differentiation, tap water rinse, TBS return blue.
10 without dehydration, direct neutral resin seal.