培養(yǎng)方法

傳代方法 將舊培養(yǎng)液吸除，PBS清洗兩遍后，加入6mL（/100mm皿）胰酶，在顯微鏡下觀察，期間禁止搖晃培養(yǎng)皿，細胞剛有脫落時，則吸除大部分胰酶，留約0.5mL，移至培養(yǎng)箱消化，約2min取出。傳代用12mL CM1-1培養(yǎng)液終止消化，輕輕吹打均勻細胞，后可分3~6皿培養(yǎng)；

生長條件 37℃，5%CO2，CM1-1培養(yǎng)液。CM1-1培養(yǎng)液：90%DMEM-H+10%FBS。DMEM-H：DMEM高糖培養(yǎng)液，含谷氨酰胺，含丙酮酸鈉。

存儲條件 凍存則用6mL凍存液（90%FBS+10%DMSO）終止消化，吹打均勻，分為6支凍存管，用程序降溫盒于-80℃凍存，過夜轉(zhuǎn)移至液氮中保存。

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培養(yǎng)操作步驟 ：

1．用蓋片鑷將蓋玻片自75%乙醇中取出，用無菌絲綢布擦拭干凈，不要用紗布； 

2．將蓋玻片輕輕放入6孔培養(yǎng)板（每孔一片）或培養(yǎng)皿中（每個平皿可放置2-3片）； 

3．在距離紫外燈直射范圍內(nèi)20-30 厘米處照射2-3小時； 

4．將經(jīng)過計數(shù)的細胞懸浮液移入培養(yǎng)板中，使蓋玻片完全浸在培養(yǎng)液中； 

5．將培養(yǎng)板在5% CO2水浴孵箱中37℃孵育2-3天，當(dāng)貼壁細胞生長至覆蓋培養(yǎng)板底部2/3面積時，將培養(yǎng)板取出，用蓋片鑷輕輕取出蓋玻片，用蒸餾水漂洗后即可進行快速固定以及細胞化學(xué)檢測。 

細胞培養(yǎng)方法：

1、細胞傳代：細胞密度達到80-90%時即可傳代

①棄去培養(yǎng)上清，用PBS或生理鹽水清洗1-2次；

②加入2ml0.25%胰酶（T25瓶），使胰酶覆蓋整個瓶或皿，蓋好放入培養(yǎng)箱消化；

③1-2min后，顯微鏡下觀察細胞，若大部分細胞回縮且有少量細胞脫落，輕輕吹打下確認(rèn)消化情況后加入完全培養(yǎng)基終止消化；若細胞還是貼壁，放回培養(yǎng)箱繼續(xù)消化至可以輕輕吹打下為止；

④將細胞懸液1000RPM左右條件下離心4min，棄上清；

⑤用新鮮培養(yǎng)基重懸后加入培養(yǎng)瓶或皿中，T25培養(yǎng)瓶加6-8ml培養(yǎng)基；

⑥懸浮細胞直接離心收集，細胞沉淀重懸后分到新培養(yǎng)瓶中。

2、細胞復(fù)蘇：

①將凍存管在37℃溫水中快速搖晃融化，時間1min左右，加入4-5ml培養(yǎng)基混勻。

②在1000RPM左右條件下離心4min，棄上清,加1-2ml培養(yǎng)基吹勻，將細胞懸液加入培養(yǎng)瓶中，補加適量培養(yǎng)基。

3、細胞凍存：待細胞生長狀態(tài)良好時進行細胞凍存保種

①棄去培養(yǎng)上清，用PBS或生理鹽水清洗1-2次，加入1mL 0.25%胰蛋白酶（T25瓶）

②1-2min后，顯微鏡下觀察細胞，大部分細胞回縮且有少量細胞脫落，輕輕吹打下確認(rèn)消化情況后加入完全培養(yǎng)基終止消化；

③將細胞懸液1000RPM左右條件下離心4min，棄上清，加1ml凍存液重懸細胞；

④將凍存管放入程序降溫盒，放入-80℃冰箱，4小時后將凍存管轉(zhuǎn)入液氮罐儲存。

 not yet tested in other applications.

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

保存條件  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.

產(chǎn)品介紹 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

英文名稱  Anti-PCDHGC5

中文名稱  原鈣粘蛋白γC5抗體

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

濃    度  1mg/1ml
H1細胞消化液（相關(guān)1）星形膠質(zhì)源性蛋白檢測試劑盒2-甲氧基-3-氯吡啶-4-酸 95% 基(2-甲基-3-呋喃基)二硫 97%

C1q腫瘤壞死因子相關(guān)蛋白6檢測試劑盒2-氯-3-甲基吡啶-5-酸 95%2-基硫 96%

核轉(zhuǎn)錄因子p65檢測試劑盒2-氯吡啶-4-酸 97%乙酸硫噻唑 99%

二酰甘油 -O-?；D(zhuǎn)移酶同源物 2檢測試劑盒5-氯噻吩-2-酸  97%2-(基硅)苯基三氟磺酸鹽 97%

胞苷脫氨酶檢測試劑盒2-氯吡啶-5-酸頻哪酯 98% 茶螺烷 technical, ≥90% (GC,mixture of isomers)

血管緊張素Ⅱ受體2檢測試劑盒2,6-二氟苯酸 98%二噻吩二硫 97%

K-Ras抗體檢測試劑盒2,6-二氟吡啶-3-酸 95%季酮酸 96%

實驗要點及說明：

1．本方法適用于貼壁細胞培養(yǎng)，而不適用于懸浮細胞培養(yǎng)，懸浮細胞可使用滴片法； 

2．所使用的蓋玻片應(yīng)該為玻璃**，并經(jīng)過鉻酸洗液處理； 

3．蓋玻片非常薄，易碎，取放蓋玻片時動作要輕； 

4．如果需要更多生長狀態(tài)一致的細胞，可以使用較大的培養(yǎng)皿，但不宜過大，以避免培養(yǎng)液的浪費和增加污染機率； 

5．如果細胞貼壁生長能力較差，可將蓋玻片在0.5%多聚賴氨酸溶液中浸泡5-10分鐘并自然晾干。