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

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甲型乙型流感診斷試劑盒（快檢方法）

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【公司名稱(chēng)】 廣州健侖生物科技有限公司
【市場(chǎng)部】    楊永漢

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【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(guò)夜
2.  二甲苯中脫蠟，梯度酒精入水(無(wú)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(kāi)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(guò)氧化氫溶液，室溫下孵育10min， 以阻斷內(nèi)源性過(guò)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?zhuān)?，陰性?duì)照采用普通血清， 室溫下孵育2小時(shí)，或4℃過(guò)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(guò)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(lái)水沖洗，蘇木素復(fù)染10min，水沖洗。0.1%的鹽酸分化，自來(lái)水沖洗，TBS返藍(lán)。
10.  不需脫水，直接用中性樹(shù)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.