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Q：Annexin V凋亡染色可以和抗体一起对细胞进行染色吗？
可以，但建议Annexin V和其它抗体分开染色。例如Annexin V、PI和CD3、CD8同时标记淋巴细胞，可以先在PBS中标记CD3/CD8后洗去，再在Binding buffer（含钙离子）中标记Annexin V。
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Q：是否有其他的检测凋亡的方法？
有的，AnnexinV检测法主要用于检测早期凋亡；中早期会出现细胞线粒体跨膜电位下降多 用JC-1等检测方法；中晚期会出现凋亡信号通路的活化，多检测Caspase3的活化水平；晚期会出现核内DNA的断裂多用Tunel方法进行检测。
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Q：Annexin VPI法为什么凋亡结果与预期不符？
处理时间过长，处理浓度过高，细胞初始活性差等原因导致的假阳性或假阴性结果。
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Q：流式测凋亡为何左上象限出奇的高？
左上象限一般是由于处理方式不当（机械力过大），处理时间太久或刺激物质浓度过高导致的环境骤变引起的非凋亡途径的细胞死亡,需要优化处理的浓度和时间梯度，以及优化细胞培养和细胞消化的流程。
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Q：Annexin VPI法如何避免出现假阳性结果？
细胞接种不易过密：接种对数生长期的细胞时密度不要＞1*106，以免细胞培养过程中自身引起凋亡，而非外界处理因素；避免消化过度：因消化过度可能会造成PS外翻，得到假阳性的结果；操作尽量轻柔：收集细胞时力度过大很可能造成细胞膜损伤，导致细胞膜的磷脂层暴露，从而结合Annexin V，导致假阳性，因此处理贴壁细胞时要小心操作，尽量避免人为的损伤。
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Q：Annexin VPI法为什么必须收集细胞上清？
因为凋亡的细胞可能会脱壁，悬浮于培养基，所以必须收集上清再离心取沉淀，合并胰酶消化下来的细胞，否则检测出来的凋亡比例可能会减少。
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Q：Annexin VPI法为什么在染色后1h内就要上机检测？
由于细胞凋亡是一个快速的过程，建议样品在染色后1小时之内进行分析；PI受时间的影响很大，因标记了PI后会加大细胞毒性，特别是检测早期凋亡时，如果时间延长除了会导致在流式细胞仪上的细胞分群差距加大外，误差会明显加大，所以建议在一个小时内检测。
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Q：What are the advantages and disadvantages of BCA protein quantification method?
advantage:
1. This method is widely used, easy to operate, fast, and can complete the test within 45 minutes. 4 times faster and even better than the classic Lowery method;
2. Not easily affected by general concentrations of detergents, urea, etc., with strong anti-interference ability;
3. The coefficient of variation for detecting different protein molecules is also much lower than that of the Coomassie Brilliant Blue method;
4. Accurate, high sensitivity, and good reagent stability. The BCA method has a good linear relationship in the concentration range of 20-20 ug/mL, and the range of trace BCA determination is 0.5-10 ug/ml.
5. Economical and practical, in addition to test tubes, measurements can be performed in microplate wells, greatly saving sample and reagent usage;
Disadvantages:
1. May be affected by chelating agents (EDTA, EGTA) and high concentrations of reducing agents (DTT, β - mercaptoethanol);
2. Incubate the reaction in a constant temperature water bath for 30 minutes or longer.
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Q：Can mycoplasma clearing agents be used together with bispecific antibodies
Do not mix with antibiotics other than streptomycin bispecific.
Li Ji Bio Quick Cell Mycoplasma Eliminator (1000X) effectively inhibits the replication of Mycoplasma DNA by suppressing the activity of Mycoplasma DNA gyrase. Starting from genetic material, it thoroughly kills Mycoplasma, allowing cells to completely eliminate the troubles of Mycoplasma contamination and ensuring the reliability of test results.
Mainly used to remove mycoplasma from cells, serum, and culture medium, the entire process only takes 3-5 days; This product can eliminate most types of mycoplasma and is non-toxic to the cells themselves.
Li Ji has validated over a hundred types of cells, such as mouse embryonic stem cells or iPS cells, human embryonic stem cells or iPS cells, HEK293, Hela, HepG2, HCT116, COS-7, Vero, Huh-7, MDCK, PANC-1, SW620, U2OS, MCF-7, MRC-5, NIH-3T3, CCC-ESF, CHO-S, CHO-K1, CHO-DG44, H295R, HL60, K562, MDA-MB-231, SP20, T47D, BM, and BV2.
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Q：Which reagent kit is better for long-term preservation of tumor tissue activity?
The live tissue cryopreservation kit vitrifies frozen tissues, effectively maintaining their morphological structure and function. The tissue activity can be efficiently preserved from the molecular level to the tissue level, achieving long-term and efficient preservation of tissue activity. It is suitable for cryopreservation of animal and tumor tissues. The live tissue resuscitation kit is used in conjunction with the cryopreservation kit for immediate resuscitation and efficient preservation of tissue activity.
Product features:
Vitrification freezing technology, no need for programmed cooling;
After recovery, the tissue activity can be preserved by more than 80%, which can be used to construct PDX/PDC models, drug sensitivity tests, and single-cell sequencing;
No bacteria, no fungi, no mycoplasma;
Sealed storage at 2-8 ℃, with a shelf life of 1 year;
"Heyuan Liji" is a joint venture between Heyuan Biotechnology (stock code: 688238) and Liji Biotechnology, specializing in the "Liji Biotechnology" and "Life-ilab" reagent brands
