H1细胞消化液（相关1）

培养方法

传代方法 将旧培养液吸除，PBS清洗两遍后，加入6mL（/100mm皿）胰酶，在显微镜下观察，期间禁止摇晃培养皿，细胞刚有脱落时，则吸除大部分胰酶，留约0.5mL，移至培养箱消化，约2min取出。传代用12mL CM1-1培养液终止消化，轻轻吹打均匀细胞，后可分3~6皿培养；

生长条件 37℃，5%CO2，CM1-1培养液。CM1-1培养液：90%DMEM-H+10%FBS。DMEM-H：DMEM高糖培养液，含谷氨酰胺，含丙酮酸钠。

存储条件 冻存则用6mL冻存液（90%FBS+10%DMSO）终止消化，吹打均匀，分为6支冻存管，用程序降温盒于-80℃冻存，过夜转移至液氮中保存。

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培养操作步骤 ：

1．用盖片镊将盖玻片自75%乙醇中取出，用无菌丝绸布擦拭干净，不要用纱布； 

2．将盖玻片轻轻放入6孔培养板（每孔一片）或培养皿中（每个平皿可放��2-3片）； 

3．在距离紫外灯直射范围内20-30 厘米处照射2-3小时； 

4．将经过计数的细胞悬浮液移入培养板中，使盖玻片完全浸在培养液中； 

5．将培养板在5% CO2水浴孵箱中37℃孵育2-3天，当贴壁细胞生长至覆盖培养板底部2/3面积时，将培养板取出，用盖片镊轻轻取出盖玻片，用蒸馏水漂洗后即可进行快速固定以及细胞化学检测。 

细胞培养方法：

1、细胞传代：细胞密度达到80-90%时即可传代

①弃去培养上清，用PBS或生理盐水清洗1-2次；

②加入2ml0.25%胰酶（T25瓶），使胰酶覆盖整个瓶或皿，盖好放入培养箱消化；

③1-2min后，显微镜下观察细胞，若大部分细胞回缩且有少量细胞脱落，轻轻吹打下确认消化情况后加入完全培养基终止消化；若细胞还是贴壁，放回培养箱继续消化至可以轻轻吹打下为止；

④将细胞悬液1000RPM左右条件下离心4min，弃上清；

⑤用新鲜培养基重悬后加入培养瓶或皿中，T25培养瓶加6-8ml培养基；

⑥悬浮细胞直接离心收集，细胞沉淀重悬后分到新培养瓶中。

2、细胞复苏：

①将冻存管在37℃温水中快速摇晃融化，时间1min左右，加入4-5ml培养基混匀。

②在1000RPM左右条件下离心4min，弃上清,加1-2ml培养基吹匀，将细胞悬液加入培养瓶中，补加适量培养基。

3、细胞冻存：待细胞生长状态良好时进行细胞冻存保种

①弃去培养上清，用PBS或生理盐水清洗1-2次，加入1mL 0.25%胰蛋白酶（T25瓶）

②1-2min后，显微镜下观察细胞，大部分细胞回缩且有少量细胞脱落，轻轻吹打下确认消化情况后加入完全培养基终止消化；

③将细胞悬液1000RPM左右条件下离心4min，弃上清，加1ml冻存液重悬细胞；

④将冻存管放入程序降温盒，放入-80℃冰箱，4小时后将冻存管转入液氮罐储存。

 not yet tested in other applications.

 optimal dilutions/concentrations should be determined by the end user.  

保存条件  Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C.

Important Note  This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.

产品介绍 The protein encoded by this gene is a histone lysine demethylase that preferentially acts on histones in the monomethyl or dimethyl states. The encoded protein requires Fe(2+) ion, 2-oxoglutarate, and oxygen for its catalytic activity. Defects in this gene are a cause of mental retardation syndromic X-linked Siderius type (MRXSSD). Four transcript variants encoding different isoforms have been found for this gene.

Function : Histone lysine demethylase with selectivity for thedi-and monomethyl states that plays a key role cell cycleprogression, rDNA transcription and brain development. Demethylatesmono- and dimethylated histone H3 'Lys-9' residue (H3K9Me1 andH3K9Me2), dimethylated H3 'Lys-27' (H3K27Me2) and monomethylatedhistone H4 'Lys-20' residue (H4K20Me1). Acts as a transcriptionactivator as H3K9Me1, H3K9Me2, H3K27Me2 and H4K20Me1 are epigeneticrepressive marks. Involved in cell cycle progression by beingrequired to control G1-S transition. Acts as a coactivator of rDNAtranscription, by activating polymerase I (pol I) mediatedtranscription of rRNA genes. Required for brain development,probably by regulating expression of neuron-specific genes. Onlyhas activity toward H4K20Me1 when nucleosome is used as a substrateand when not histone octamer is used as substrate. May also haveweak activity toward dimethylated H3 'Lys-36' (H3K36Me2), however,the relevance of this result remains unsure in vivo. Specificallybinds trimethylated 'Lys-4' of histone H3 (H3K4me3), affectinghistone demethylase specificity: has weak activity toward H3K9Me2in absence of H3K4me3, while it has high activity toward H3K9me2when binding H3K4me3.

Subunit : Interacts with POLR1B, UBTF, SETD1A, HCFC1, E2F1 andZNF711.

Subcellular Location : Nucleus. Nucleus, nucleolus. Note=Recruitedto H3K4me3 sites on chromatin during interphase. Dissociates fromchromatin when cells enter mitosis.

Post-translational modifications : Phosphorylation at Ser-69 and Ser-120 are required fordissociation from chromatin and accumulation of H4K20Me1 levelsduring prophase.

DISEASE : Defects in PHF8 are the cause of mental retardationsyndromic X-linked Siderius type (MRXSSD) [MIM:300263]. A disordercharacterized by mild to borderline mental retardation with orwithout cleft lip/cleft palate.

Similarity : Belongs to the JHDM1 histone demethylase family.JHDM1D subfamily.

Contains 1 JmjC domain.

Contains 1 PHD-type zinc finger.

Database links : UniProtKB/Swiss-Prot: Q9UPP1.3

英文名称  Anti-PCDHGC5

中文名称  原钙粘蛋白γC5抗体

别    名  PCDH-gamma-C5; Protocadherin gamma C5; protocadherin gamma subfamily C, 5; MGC138286; MGC138288; PCDGM_HUMAN.

浓    度  1mg/1ml
H1细胞消化液（相关1）星形胶质源性蛋白检测试剂盒2-甲氧基-3-氯吡啶-4-酸 95% 基(2-甲基-3-呋喃基)二硫 97%

C1q肿瘤坏死因子相关蛋白6检测试剂盒2-氯-3-甲基吡啶-5-酸 95%2-基硫 96%

核转录因子p65检测试剂盒2-氯吡啶-4-酸 97%乙酸硫噻唑 99%

二酰甘油 -O-酰基转移酶同源物 2检测试剂盒5-氯噻吩-2-酸  97%2-(基硅)苯基三氟磺酸盐 97%

胞苷脱氨酶检测试剂盒2-氯吡啶-5-酸频哪酯 98% 茶螺烷 technical, ≥90% (GC,mixture of isomers)

血管紧张素Ⅱ受体2检测试剂盒2,6-二氟苯酸 98%二噻吩二硫 97%

K-Ras抗体检测试剂盒2,6-二氟吡啶-3-酸 95%季酮酸 96%

实验要点及说明：

1．本方法适用于贴壁细胞培养，而不适用于悬浮细胞培养，悬浮细胞可使用滴片法； 

2．所使用的盖玻片应该为玻璃**，并经过铬酸洗液处理； 

3．盖玻片非常薄，易碎，取放盖玻片时动作要轻； 

4．如果需要更多生长状态一致的细胞，可以使用较大的培养皿，但不宜过大，以避免培养液的浪费和增加污染机率； 

5．如果细胞贴壁生长能力较差，可将盖玻片在0.5%多聚赖氨酸溶液中浸泡5-10分钟并自然晾干。