Hybridoma Cell Expansion And Subcloning Medium 杂交瘤细胞扩培及亚克隆专用培养基

产品信息 Product description

产品名称

Product Name

杂交瘤细胞扩培及亚克隆专用培养基

Hybridoma Cell Expansion and Subcloning Medium

货号 Cat.No

BRS-MED02

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)

Fetal Bovine Serum (FBS, imported): 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

其它添加物：单细胞增殖因子及小分子添加剂

Other Additives: Single-cell proliferation factors and small molecule 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 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, then collect the supernatant for use.

融合孔处理(状态良好细胞):

针对状态良好的融合孔(比如孔内培养基颜色正常或轻微发黄,且显微镜下细胞状态良好,透亮,大小正常等)可直接使用黄枪头轻轻吹打融合孔,细胞计数后按0.8-1.5cell/孔/200ul培养基铺板.

Handling Fusion Wells – Healthy Cells:

For fusion wells with cells in good condition (e.g., medium color normal or slightly yellow, cells under microscope appear healthy, bright, and normal in size), gently pipette up and down with a yellow pipette tip. Count the cells and plate at 0.8–1.5 cells/well in 200 µL medium.

融合孔处理(状态欠佳细胞):

针对状态不太良好的融合孔(比如孔内培养基颜色酸化发黄或显微镜下细胞状态皱缩,发黑等)可选择两种方式进行后续亚克隆.

Handling Fusion Wells – Suboptimal Cells:

For fusion wells with cells in suboptimal condition (e.g., medium acidified/yellow or cells appear shriveled, dark under microscope), two options are available for subsequent subcloning:

第一种方式是将融合孔细胞吸至装有1ml本培养基的24孔板内,在扩培后的第2-3天再行计数进行亚克隆铺板(0.8-1.5cell/孔/200ul培养基),此时融合孔细胞倍增的次数有限且细胞状态良好,非常适于亚克隆铺板;

Option 1: Transfer cells from the fusion well into a 24-well plate containing 1 mL of this medium. After 2–3 days of expansion, count the cells and perform subcloning at 0.8–1.5 cells/well in 200 µL medium. At this stage, cell doubling is limited but cell condition is good, making it ideal for subcloning.

第二种方式是使用黄枪头小心吸弃融合孔上清,每孔补充200ul新鲜的本培养基,至37度5% CO2静置培养3-4小时,然后取样细胞计数后进行亚克隆(0.8-1.5cell/孔/200ul培养基);

Carefully remove the supernatant from the fusion well using a yellow pipette tip and add 200 µL of fresh medium per well. Incubate at 37°C, 5% CO₂ for 3–4 hours. Then, sample the cells, count, and perform subcloning at 0.8–1.5 cells/well in 200 µL medium.

注：使用本司"杂交瘤扩培及亚克隆专用培养基"培养后的亚克隆一般第6-8天即可开始下游检测筛选.

Note: Subclones cultured with the Hybridoma Cell Expansion and Subcloning Medium typically reach a stage suitable for downstream screening or analysis on days 6–8 post-subcloning.