145-2C11(仓鼠/小鼠杂交瘤细胞(产生CD3单抗))General information
Synonyms:145-2C11
Background:Animals were immunized with the BM10-37 mouse cytotoxic T lymphocyte (CTL) cell clone (anti H-2 Kb). Spleen cells were fused with Sp2/0-Ag14 myeloma cells.
Species:Cricetulus migratorius (B cell); Mus musculus (myeloma), hamster, Armenian (B cell); mouse (myeloma)
Tissue:Spleen, blood
Morphology:Lymphoblast-like
Growth Mode:Suspension
Culture Medium:IMDM+10%FBS
We strongly suggest to purchase the complete medium from Cobioer.
Cryopreservation medium:90%FBS+10%DMSO
Genes Expressed:immunoglobulin; monoclonal antibody; against mouse CD3
Cellular Products:immunoglobulin; monoclonal antibody; against mouse CD3
细胞冷冻保存:
1.冻存液:92%完全培养基+8%DMSO(可以根据实验室条件自行选择)
2.降温步骤:4度10min,-20度2h,-80过夜后液氮保存。
细胞培养的操作步骤:
1.145-2C11(仓鼠/小鼠杂交瘤细胞(产生CD3单抗))吸走用于培养基,加入3-5mL PBS后轻轻晃动培养瓶润洗细胞;
2.吸干净PBS后加入1mL 0.25%胰酶-0.53mM EDTA,轻轻摇晃培养皿使胰酶浸没细胞表面,培养瓶放37度培养箱消化;
(细胞对于消化比较敏感,消化过度会严重影响细胞状态,导致细胞传代死亡漂浮或者传代后不长。细胞消化到细胞间隙变大但未脱落,可以轻轻吹下时即可终止,禁止消化到细胞完全漂浮。混匀细胞时尽量轻柔,不要吹出大量气泡。)
3.加入6-8ml完全培养基终止消化,轻轻吹下细胞混匀;
4.将混匀的细胞1000rpm(约150g)离心3min,弃上清,再用新鲜培养基重悬37度培养箱继续培养;
传代比例: 不同细胞生长速度不一,具体传代比例视细胞生长速度而定,大部分细胞适用1:3-1:4传代,生长较慢的细胞可以1:2传代。