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乙型流感病毒抗原檢測(cè)試劑盒(免疫層析法)
廣州健侖生物科技有限公司
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乙型流感病毒抗原檢測(cè)試劑盒(免疫層析法)
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【公司名稱】 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
3,產(chǎn)生組織切片非特異性染色
1)抗體孵育時(shí)間過(guò)長(zhǎng)、抗體濃度高易增加背景著色。這可通過(guò)縮短一抗/二抗孵育時(shí)間、稀釋抗體來(lái)控制。這是zui重要的一條;
2)一抗用多克隆抗體易出現(xiàn)非特異性著色,建議試用單克隆抗體看看;
3)內(nèi)源性過(guò)氧化物酶和生物素在肝臟、腎臟等組織含量很高(含血細(xì)胞多的組織),需要通過(guò)延長(zhǎng)滅活時(shí)間和增加滅活劑濃度來(lái)降低背景染色;
4)非特異性組分與抗體結(jié)合,這需要通過(guò)延長(zhǎng)二抗來(lái)源的動(dòng)物免疫血清封閉時(shí)間和適當(dāng)增加濃度來(lái)加強(qiáng)封閉效果;
5)DAB孵育時(shí)間過(guò)長(zhǎng)或濃度過(guò)高;
6)PBS沖洗不充分,殘留抗體結(jié)果增強(qiáng)著色,在一抗、二抗或SP孵育之后的浸洗尤為重要;
7)標(biāo)本染色過(guò)程中經(jīng)常出現(xiàn)干片,這容易增強(qiáng)非特異性著色。
4,免疫組化染色呈陰性結(jié)果
1)抗體濃度和質(zhì)量問(wèn)題以及抗體來(lái)源選擇錯(cuò)誤;
2)抗原修復(fù)不全,對(duì)于甲醛固定的組織必須用充分抗原修復(fù)來(lái)打開(kāi)抗原表位,以利于與抗體結(jié)合;建議微波修復(fù)用高火4次*6min試試。有人做過(guò)實(shí)驗(yàn),這是*的時(shí)間和次數(shù)。若不行,還可高壓修復(fù);
3)組織切片本身這種抗原含量低;
4)血清封閉時(shí)間過(guò)長(zhǎng);
5)DAB孵育時(shí)間過(guò)短;
6)細(xì)胞通透不全,抗體未能充分進(jìn)入胞內(nèi)參與反應(yīng);
7)開(kāi)始做免疫組化,我建議你一定要首先做個(gè)陽(yáng)性對(duì)照片,排除抗體等外的方法問(wèn)題。
3, resulting in non-specific tissue section staining
1) antibody incubation time is too long, high antibody concentration easily increase background coloring. This can be controlled by reducing the primary antibody / secondary antibody incubation time and diluting the antibody. This is the most important one
2) The first antibody polyclonal antibody prone to non-specific coloring, it is recommended to try monoclonal antibody to see;
3) endogenous peroxidase and biotin in the liver, kidneys and other tissues is very high (including blood cells and more tissue), need to extend the inactivation time and increase the concentration of inactivator to reduce the background staining;
4) non-specific components and antibodies, which need to extend the secondary antibody from the immune serum to close time and the appropriate increase in concentration to enhance the sealing effect;
5) DAB incubation time is too long or too high concentration;
6) Inadequate washing with PBS, enhanced staining with residual antibody results, and dipping after primary antibody, secondary antibody or SP incubation are particularly important;
7) Dry specimens often appear during specimen dyeing, which easily enhances nonspecific staining.
4, immunohistochemistry showed negative results
1) Antibody concentration and quality problems and wrong selection of antibody sources;
2) incomplete antigen repair, fixed tissue for formaldehyde must be fully antigen retrieval to open the antigen epitope, in order to facilitate the combination of antibodies; recommended microwave repair with high fire 4 * 6min try. Someone has done experiments, this is the best time and frequency. If not, but also high-pressure repair;
3) the tissue section itself is low in this antigen;
4) serum blocking time is too long;
5) DAB incubation time is too short;
6) cells are not fully permeable, the antibody failed to fully enter the cell involved in the reaction;
7) begin to do immunohistochemistry, I suggest that you must first be a positive pair of photos, excluding methods other than antibodies.