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。
产品名称
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规格
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货号
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鸡胚胎成纤维细胞
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1×106
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BH-X019753
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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;部分产品现货,超低比价,货期短,价格优,公司产品仅用于科研售后齐全。
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(每个平皿可放置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。
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 Pbx 1, 2, 3 and 4 are members of the TALE (three amino acid loop extension) family of homeodomain-containing proteins. Human pre-B cell acute leukemias are frequently associated with a t(1;19)(q23;p13.3) chromosomal rearrangement, which creates a chimeric gene encoding a fusion between the E2A and Pbx 1 gene products. Pbx 2 and Pbx 3 share 92% and 94% respective identities with Pbx 1 over a 266 amino acid region flanking their homeobox domains, while all three proteins are quite divergent at their amino- and carboxy-termini. Two forms of Pbx 1 and Pbx 3 each differ primarily in their carboxy-termini and result from alternative mRNA splicing. Unlike other homeotic selector genes which are expressed transiently during development and differentiation, Pbx gene transcripts are ubiquitously expressed in both fetal and ***** tissues and cell lines. Additionally, Pbx 2 and Pbx 3 transcripts are detected in lymphoid cells, which do not express Pbx 1. Pbx 4 expression is confined to the testis, especially to spermatocytes in the pachytene stage of the first meiotic prophase.
Subcellular Location : Nucleus.
Similarity : Belongs to the TALE/PBX homeobox family.
Contains 1 homeobox DNA-binding domain.
Database links : UniProtKB/Swiss-Prot: Q9BYU1.2
英文名称 Anti-PHEX
中文名称 维生素D低磷性佝偻病蛋白
别 名 HPDR; HPDR1; HYP; HYP1; LXHR; Metalloendopeptidase homolog PEX; PEX; Phex; PHEX_HUMAN; Phosphate regulating endopeptidase homolog X linked; Phosphate regulating gene with homologies to endopeptidases on the X chromosome; Phosphate-regulating neutral endopeptidase; Vitamin D-resistant hypophosphatemic rickets protein; X-linked hypophosphatemia protein; XLH.
浓 度 1mg/1ml
规 格 0.2ml/200μg
抗体来源 Rabbit
克隆类型 polyclonal
交叉反应 Human, Mouse, Rat, Dog, Pig, Cow, Horse, Rabbit, Sheep
产品类型 一抗
研究领域 发育生物学 干细胞
蛋白分子量 predicted molecular weight: 86kDa
鸡胚胎成纤维细胞髓样分化蛋白2检测试剂盒次氯酸钠溶液 AR,6-14% active chlorine basisL-(-)-二(对甲酰) 97%
羧化不全骨钙素检测试剂盒次氯酸钠 CP,available chlorine 5.5 %3-(二甲氨基)烯酸乙酯 98%
羧甲基赖氨酸检测试剂盒次氯酸钠溶液 ACS,available chlorine 5.00 %3-氨基巴豆酸乙酯 99%
羧酸酯酶检测试剂盒硫代乙酸钠 AR,80.0%硫代草氨酸乙酯 95%
缩酶A检测试剂盒硫代乙酸钠 BR乙酸乙酯 98%
他克莫司检测试剂盒硫代乙酸钠 98%二乙酯 97%
胎儿纤维连接蛋白检测试剂盒亚硝基铁化钠 AR,99.0%3-(5-乙基-1,2,4-噁二唑-3-基)苯甲酸 97%
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。