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產(chǎn)品名稱
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規(guī)格
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貨號
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倉鼠卵巢細胞
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1×106
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BH-X019738
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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。
培養(yǎng)操作步驟 :
1.用蓋片鑷將蓋玻片自75%乙??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檢測。

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。
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.
phosphorylating the RELA subunit at 'Ser-311'. Is necessary and sufficient for LTP maintenance in hippocampal CA1 pyramidal cells.
Subunit : Forms a ternary complex with SQSTM1 and KCNAB2. Forms another ternary complex with SQSTM1 and GABRR3. Forms a complex with SQSTM1 and MAP2K5 (By similarity). Interacts with PARD6A, PARD6B, PARD6G and SQSTM1. Part of a complex with PARD3, PARD6A or PARD6B or PARD6G and CDC42 or RAC1. Interacts with ADAP1/CENTA1. Forms a ternary complex composed of SQSTM1 and PAWR. Interacts directly with SQSTM1 (Probable). Interacts with IKBKB. Interacts (via the protein kinase domain) with WWC1. Forms a tripartite complex with WWC1 and DDR1, but predominantly in the absence of collagen. Component of the Par polarity complex, composed of at least phosphorylated PRKCZ, PARD3 and TIAM1. Interacts with PDPK1 (via N-terminus region).
Subcellular Location : Cytoplasm. Endosome. Cell junction. Note=In the retina, localizes in the terminals of the rod bipolar cells. Associates with endosomes. Presence of KRIT1, CDH5 and RAP1B is required for its localization to the cell junction.
Tissue Specificity : Expressed in brain, and to a lesser extent in lung, kidney and testis.
Post-translational modifications : CDH5 is required for its phosphorylation at Thr-410. Phosphorylated by protein kinase PDPK1; phosphorylation is inhibited by the apoptotic C-terminus cleavage product of PKN2. Phosphorylation at Thr-410 by PI3K activates the kinase.
Similarity : Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PKC subfamily.
Contains 1 AGC-kinase C-terminal domain.
Contains 1 OPR domain.
Contains 1 phorbol-ester/DAG-type zinc finger.
Contains 1 protein kinase domain.
Database links : UniProtKB/Swiss-Prot: Q05513.4
英文名稱 Anti-PIM1+PIM3
中文名稱 絲氨酸/蘇氨酸激酶蛋白PIM1+PIM3抗體
倉鼠卵巢細胞狀腺原氨酸檢測試劑盒2,5-二氯噻吩 98%氫氧化鋁 99.99%,2~10μm
氧芐氨嘧啶檢測試劑盒3,4-二甲氧基噻吩 97%氫氧化鋁 99.99%,0.05~1.5μm
三磷酸磷脂酰肌檢測試劑盒3-甲氧基噻吩 98%納米氧化鎂 50nm,球形,99.9% metals basis
三磷酸腺苷檢測試劑盒2-甲氧基噻吩 99%二氧化鋯 99.99% metals basis,D50:0.2~0.4μm
三葉因子3檢測試劑盒2-硝基噻吩 98%納米氧化鋅 99.9% metals basis,30±10nm
色氨酰tRNA合成酶檢測試劑盒四溴噻吩 99%5-氨基乳清酸 98%
色素上皮衍生因子檢測試劑盒噻吩-2,3-二甲 98%1,3-二甲基-5-吡唑酮 96%