實(shí)驗(yàn)要點(diǎn)及說明：
1．本方法適用于貼壁細(xì)胞培養(yǎng)，而不適用于懸浮細(xì)胞培養(yǎng)，懸浮細(xì)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(shí)動(dòng)作要輕；
4．如果需要更多生長(zhǎng)狀態(tài)一致的細(xì)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(fèi)和增加污染機(jī)率；
5．如果細(xì)胞貼壁生長(zhǎng)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。
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產(chǎn)品名稱
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規(guī)格
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貨號(hào)
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原代前脂肪細(xì)胞培養(yǎng)體系
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100ml/500ml
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BH-X019968
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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(xì)胞剛有脫落時(shí)，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(xì)胞，后可分3~6皿培養(yǎng)；
生長(zhǎng)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(chǔ)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。
我司常期代理ATCC Acris abcam cst Biorbyt santa Novus sigma lifespan NEB roche ABI R&D millipore BD Qiagen Cayman Jackson Life GeneTex Bio-Rad DSHB tocris peprotech 等品牌；部分產(chǎn)品現(xiàn)貨，超低比價(jià)，貨期短，價(jià)格優(yōu)，公司產(chǎn)品僅用于科研售后齊全。
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(gè)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(shí)；
4．將經(jīng)過計(jì)數(shù)的細(xì)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(xì)胞生長(zhǎng)至覆蓋培養(yǎng)板底部2/3面積時(shí)，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(jìn)行快速固定以及細(xì)胞化學(xué)檢測(cè)。
細(xì)胞培養(yǎng)方法：
1、細(xì)胞傳代：細(xì)胞密度達(dá)到80-90%時(shí)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(gè)瓶或皿，蓋好放入培養(yǎng)箱消化；
③1-2min后，顯微鏡下觀察細(xì)胞，若大部分細(xì)胞回縮且有少量細(xì)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(xì)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(xì)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(xì)胞直接離心收集，細(xì)胞沉淀重懸后分到新培養(yǎng)瓶中。
2、細(xì)胞復(fù)蘇：
①將凍存管在37℃溫水中快速搖晃融化，時(shí)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(xì)胞懸液加入培養(yǎng)瓶中，補(bǔ)加適量培養(yǎng)基。
3、細(xì)胞凍存：待細(xì)胞生長(zhǎng)狀態(tài)良好時(shí)進(jìn)行細(xì)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(xì)胞，大部分細(xì)胞回縮且有少量細(xì)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(xì)胞懸液1000RPM左右條件下離心4min，棄上清，加1ml凍存液重懸細(xì)胞；
④將凍存管放入程序降溫盒，放入-80℃冰箱，4小時(shí)后將凍存管轉(zhuǎn)入液氮罐儲(chǔ)存。
亞 型 IgG
純化方法 affinity purified by Protein A
儲(chǔ) 存 液 0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide
產(chǎn)品應(yīng)用 WB=1:100-500 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 IF=1:100-500
（石蠟切片需做抗原修復(fù)）
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 HILI is a 973 amino acid protein encoded by the human gene PIWIL2. HILI belongs to the argonaute family and contains one PAZ domain and one PIWI domain. HILI is a cytoplasmic protein that is expressed in ***** testis and in most tumors. It regulates spermatogenesis and primordial germ cell production and has an essential role in meiotic differentiation of spermatocytes and in self-renewal of spermatogonial stem cells. Expression of HILI can modulate expression of genes involved in stem cell proliferation (such as PDGFR-b), in energy metabolism (such as Glut1), in cell-cell interaction (such as Integrin a6, GJA7, THY-1 and CD9), and in germ cell differentiation (such as STRA8). It may also play a role as a regulatory factor of Stat3/Bcl-xS/L/CCND1 pathway. Repression of HILI can inhibit tumor cell growth. HILI acts as an oncogene by inhibition of apoptosis and promotion of proliferation in tumors.
CD9 and STRA8. Inhibits tumor cell growth when repressed. When overexpressed, acts as an oncogene by inhibition of apoptosis and promotion of proliferation in tumors.
Subunit : Interacts with DDX4, MAEL, EIF3A, EIF4E, EIF4G, PRMT5 and WDR77. Associates with EIF4E- and EIF4G-containing m7G cap-binding complexes. Interacts (when methylated on arginine residues) with TDRD1.
Subcellular Location : Cytoplasm. Note=Present in chromatoid body. Probable component of the meiotic nuage, also named P granule, a germ-cell-specific organelle required to repress transposon during meiosis.
Tissue Specificity : Expressed in ***** testis and in most tumors.
Post-translational modifications : Arginine methylation by PRMT5 is required for the interaction with Tudor domain-containing protein TDRD1 and subsequent localization to the meiotic nuage, also named P granule.
Similarity : Belongs to the argonaute family. Piwi subfamily.
Contains 1 PAZ domain.
Contains 1 Piwi domain.
原代前脂肪細(xì)胞培養(yǎng)體系解偶聯(lián)蛋白 1檢測(cè)試劑盒4-溴-4',4''-二甲基三苯 95%對(duì)甲 98%
her2-ecd檢測(cè)試劑盒聯(lián)芐 99%對(duì)氨基酚 CP，98%
分泌型卷曲相關(guān)蛋白3檢測(cè)試劑盒2-叔丁基蒽醌 98%對(duì)氨基酚 99% (HPLC)
鳥氨酸檢測(cè)試劑盒4,4'-聯(lián)苯二甲酸 97%間氯 CP,98%
肽基精氨酸脫亞酶2檢測(cè)試劑盒4,4'-二甲基二苯 97%間氯 GR，99%
肽基精氨酸脫亞酶4檢測(cè)試劑盒3,4-二甲基二苯 98%鄰氯 GR,99%
結(jié)核桿菌抗體檢測(cè)試劑盒4,4'-二甲氧基二苯基 98%鄰氯 GCS，>99.5% (GC)