6、Agrisera植物一抗來源物種對WB實驗是否有影響？是否起重要作用？
7、如有Agrisera植物一抗可以應用于immunolocalization是否也能應用于WB？
8、如有Agrisera植物一抗可以適用于擬南芥，是否也能適用于其他物種？
9、如某一Agrisera植物一抗產(chǎn)品說明書中未提到客戶待檢測物種，應該如何判斷此一抗是否能適用于此待檢測物種？
10、客戶購買Agrisera抗體，使用后實驗結果不理想，怎么辦？
12、Agrisera未上架的抗體申請試用樣品，需要客戶提供哪些信息？
1、Agrisera的抗體是否可以提供試用樣品？
在售產(chǎn)品不提供試用樣品。未上架產(chǎn)品需要客戶提供非常詳細的實驗樣品信息后由Agrisera廠家評估發(fā)放，而且需要客戶在收到試用樣品后三個月內(nèi)提供實驗結果。
2、Agrisera等待上架產(chǎn)品預計供貨時間？
由于植物抗體驗證需要各類樣本，突變體對照等，樣品準備周期很長，所以導致一般植物抗體驗證周期非常長，1-2年時間都會出現(xiàn)。
4、Agrisera抗體在血清的濃度是多少？
大部分情況，特異性抗體在血清的濃度在0.05-0.2mg/ml之間。特殊情況下，表達強的多抗血清中包含有1-3mg/ml的特異性抗體，甚至更多。
5、應該如何保存Agrisera抗體？能保存多久？
抗體盡量避免反復凍融，收到抗體后根據(jù)實驗需要對抗體進行分裝。嚴格按照說明書推薦的保存條件正確處理和保存抗體。
Each antibody is different, some might last for 20-40 years, while other antibodies will have quite a short life of one year. Storage conditions are of importance to extend antibody life, however the stability of each specific antibody might be different. Therefore, please follow the information provided on the specific product information sheet for each antibody. What works for one antibody, might not necessarily be applicable for another one. This is very important to keep in mind. If it is not clear ，pleasecontact us. General recommendations for antibody storage can be found here.
6、Agrisera植物一抗來源物種對WB實驗是否有影響？是否起重要作用？
It does not matter from which species the primary antibody comes from for a western blot application. There are many good secondary antibodies available for any of those species you inquire about.
What is important is to which part of a protein an antibody is made to. Is it going to recognize epitopes (stretches of 2-3 amino acids) which are exposed on a given protein after transfer to a membrane? Some proteins can refold following the transfer, depending upon applied conditions, which can make a given epitope not accessible. It also depends on if the western blot is performed in native or denatured conditions. The addition of 6-8 M urea to your loading buffer might help to keep your protein unfolded and accessible for an antibody to bind.
Before you use an antibody in western blot, please check to which part of the protein it was made. Are there any references which confirms the use of this antibody for this specific technique?
7、如有Agrisera植物一抗可以應用于immunolocalization是否也能應用于WB？
不一定
Again, it depends upon the antibody binding place on the protein. Is this part exposed after fixation or following the western blot? If a polyclonal antibody is made to a synthetic peptide, it is recognizing a pool of epitopes (linear epitopes are streches of 2-3 amino acids)and if such epitopes are not exposed following fixation: the antibody is not going to work.
8、如有Agrisera植物一抗可以適用于擬南芥，是否也能適用于其他物種？
不一定，具體適用物種請查看產(chǎn)品說明書。
Not really, it all depends upon the conservation level of a specific peptide used to elicit this antibody. It can be checked by comparing the peptide used to elicit this antibody and your protein sequence. The conservation level between the peptide used to make an antibody and the peptide found in your protein seqence needs to be around 80 %, to allow an antibody to work in case of anti-peptide antibodies and can be lower for antibodies made to larger parts of a protein. In some cases 6-8 M urea needs to be included to fully unfold the protein. The electrophoretic mobility of a protein can also vary between different species.
9、如某一Agrisera植物一抗產(chǎn)品說明書中未提到客戶待檢測物種，應該如何判斷此一抗是否能適用于此待檢測物種？
需要客戶提供待檢測物種的特定蛋白序列，Agrisera廠家根據(jù)客戶提供的蛋白序列進行蛋白比對，評估是否可以適用于待檢測物種。
10、客戶購買Agrisera抗體，使用后實驗結果不理想，怎么辦？
需要客戶提供具體實驗信息，填寫使用反饋表，Agrisera廠家會根據(jù)客戶反饋情況提供具體優(yōu)化方案。
Have you checked recommended conditions to use for this specific antibody, included on the product info sheet? Some antibodies will work reasonably well and give good results regardless of the applied conditions, like membrane type, load per well and developing reagent. Some antibodies will actually require a bit more consideration. Less backgrond can be obtained for some antibodies when using pvdf vs. nitrocellulose membrane for transfer or by applying recommended blocking. There are also variations between secondary antibodies. At least, but the most important, is the type of sample you are analyzing. You can eithercontact us or check our Western blot trouble shooting, it can be found here.
11、客戶如何選擇合適的Agrisera植物抗體？
Usually the C or N-terminal of the protein is used, as those parts of protein are exposed. Also, to mimic protein behavior, the synthesized peptide should have similar structure and charge as the protein it has been "cut out off". Therefore:
- peptides derived from the C terminal should have N terminal modified by acetylation
- peptides derived from the N terminal should have C terminal modified by amidation - peptides derived from an internal sequence should have both ends modified
Following points should also be considered:
- Are there any other proteins from the family of interest, where cross-reactivity should be avoided?
- Is the crystal structure of the protein (or homologous protein) known? This would be helpful for the peptide chemist in searching for the best peptide for antibody production.
- What is the final application of the produced antibodies? Native or denatured techniques?
12、Agrisera未上架的抗體申請試用樣品，需要客戶提供哪些信息？
檢測物種、實驗類型、蛋白序列，客戶能否承諾收到試用樣品后三個月內(nèi)給予實驗反饋。Agrisera廠家根據(jù)客戶提供的信息評估發(fā)放試用樣品。
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