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杂交瘤细胞的作育,融入与克隆皇家赌场号hj85:

文章作者:生命科学 上传时间:2019-08-27

着力提醒:Hancock Laboratory Methods,Department of Microbiology and Immunology,University of British Columbia, British Columbia

骨干提示:Reagents Medium A - Pre-fusion Medium and Hybridoma Expansion Medium

主干提醒:Materials Tumor cells that have been treated with 8-azaguanine for 48 hours are removed from the drug and grown to Materials

Hancock Laboratory Methods,Department of Microbiology and Immunology,University of British Columbia, British Columbia, Canada

  1. Medium A - Pre-fusion Medium and Hybridoma Expansion Medium
  2. Medium B - Fusion Medium
  3. Medium C - Hybridoma Recovery Medium
  4. Medium D - Hybridoma Selection Medium
  5. Medium E - Hybridoma Growth Medium
  6. PEG Solution
  1. Tumor cells that have been treated with 8-azaguanine for 48 hours are removed from the drug and grown to a maximum concentration of 500,000 cells per ml.
  2. Rats or mice that were previously immunized and then boosted IV 72 hours prior to hybridization.
  3. Media: Auto-Pow with sodium bicarbonate plus non-essential amino acids, penicillin-streptomycin, L-glutamine and HT. For the serum-containing media , add 5 - 10% newborn calf serum.
  4. 40% PEG: Aldrich 1000. Made up in serum-free medium . The stock may be aliquoted and stored at -30°C.
  5. Other materials include: 96-well plates, sterile Petri dishes, conical 15 and 50 ml tubes, round bottom tubes, sterile dissection instruments, 37°C water bath, spleen crusher, 70% ethanol, syringe and needle.

Materials

Procedure Day 0

Kill immunized mouse by CO2 asphyxia and wet throughly with ethanol. Aseptically remove spleen and place on sterile cell sieve . Cut the spleen into small pieces with sterile scissors and press through the screen with the end of the plunger of a 1ml sterile syringe into a sterile petri dish containing 10ml of DMEM-10

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