Hybridoma Fusion And Selection Medium (HAT) 杂交瘤细胞融合筛选培养基（HAT）

产品信息 Product description

产品名称

Product Name

杂交瘤细胞融合筛选培养基(HAT)

Hybridoma Fusion and Selection Medium (HAT)

货号 Cat.No

BRS-MED01

规格

Specification

500ml/瓶

500 mL / bottle

产品组分

Components

DMEM 基础培养基：80%(体积比)

DMEM Base Medium: 80% (v/v)

进口胎牛血清：10%(体积比)

Imported Fetal Bovine Serum (FBS): 10% (v/v)

条件培养基：10%(体积比)

Conditioned Medium: 10% (v/v)

青霉素：100 单位/mL

Penicillin: 100 U/mL

链霉素：100 单位/mL

Streptomycin: 100 U/mL

次黄嘌呤(Hypoxanthine)：100uM

Hypoxanthine: 100 µM

胸腺嘧啶核苷(Thymidine)：16uM

Thymidine: 16 µM

氨基蝶呤(Aminopterin)：0.4uM

Aminopterin: 0.4 µM

其它添加物：多种细胞因子(足量)及添加剂

Other Additives: Various cytokines (sufficient amount) and supplements

保存条件

Storage conditions

-20℃保存,有效期 18 个月.

Store at -20°C. Shelf life: 18 months.

使用方法

Instructions for Use

培养基融化：

37℃水浴融化本培养基,乙醇消毒外表面后置超净台备用;

注：本品融化后出现少量白色絮状物属正常现象,静置 3-5min 使其沉至瓶底,然后吸取上清使用即可.

Thawing the Medium:

Thaw the medium in a 37°C water bath. Wipe the outer surface with 70% ethanol and place it in a biosafety cabinet for use.
Note: The appearance of a small amount of white flocculent material after thawing is normal. Allow the medium to stand for 3–5 minutes for the flocs to settle at the bottom, then carefully collect the supernatant for use.

细胞融合与接种:

使用物理(电融合)或化学(常规PEG)的方法完成细胞融合,然后使用本培养基直接重悬融合细胞,以合适的密度(电融合推荐：1E4 细胞/孔-2E4 细胞/孔;PEG 融合推荐：5E4 细胞/孔-1E5 细胞/孔)和体积 (推荐：200ul/孔)铺 96 孔板,置 37℃ 5% CO2 培养箱静置培养.

Cell Fusion and Seeding:

Perform cell fusion using physical (electrofusion) or chemical (conventional PEG) methods. Resuspend the fused cells directly in this medium at an appropriate density and volume:

Electrofusion: 1 × 10⁴ – 2 × 10⁴ cells/well

PEG fusion: 5 × 10⁴ – 1 × 10⁵ cells/well
Recommended volume: 200 µL/well in a 96-well plate.
Incubate at 37°C, 5% CO₂ for culture.

培养液更换

融合后的第 4-8 天,根据细胞生长状态进行半换液或全换液培养;

Medium Replacement:

On days 4–8 post-fusion, perform partial or complete medium replacement according to the growth status of the cells.

克隆筛选：

当孔中细胞克隆长至合适大小后进行融合孔阳筛检测;

Clone Screening:

When the cell clones in the wells reach an appropriate size, perform positive clone screening.

注意事项

Precautions

本产品有一定毒性,仅用于杂交瘤细胞的筛选及培养,不可用于 培养活体动物,诊断或治疗药物用.

This product is toxic and is intended only for the selection and culture of hybridoma cells. It must not be used for the culture of live animals, diagnostic purposes, or therapeutic applications.

本产品适用于 HGPRT 筛选系统的杂交瘤细胞的筛选培养,不适用于 TK 或其它选择系统的细胞培养.本品不能用于直接培养 HGPRT 缺陷细胞 (如 NS1, SP2/0 等).

This product is suitable for the selection and culture of hybridoma cells using the HGPRT selection system. It is not applicable for TK or other selection systems. This medium cannot be used for direct culture of HGPRT-deficient cells (e.g., NS1, SP2/0).

本产品为无菌溶液,pH 为 7.2~7.4,取用需在无菌条件下进行.

This product is a sterile solution with a pH of 7.2–7.4. All handling must be performed under aseptic conditions.