Editing Antibody Production in China (section)

== Materials ==

*Up to ten 6-8 weeks old female Balb/c&nbsp; mice from Wushi Animal Laboratory (Shanghai, China).
*QuickAntibody-Mouse2W（2周快速鼠多抗制备佐剂), 3 ml, KX0210043, 600Yuan/ml) from 北京博奥龙免疫技术有限公司[http://en.biodragon.cn/m/list.php?tid=27 http://en.biodragon.cn/m/list.php?tid=27]
*SP2/0 myeloma cells
*Nunc MaxiSorp™ flat-bottom from ThermoFisher
*HRP-labeled goat anti-mouse IgG
*HRP substrate
*2 M H2SO4
*Microplate Reader with 450 nm

Sigma-Aldrich (Shanghai, China):&nbsp;<br/>Hypoxanthine, aminopterin and thymidine supplement (HAT), hypoxanthine and thymidine supplement (HT)&nbsp;

Thermo Fisher Scientific (Shanghai, China): RPMI 1640 medium powder,&nbsp; fetal bovine serum (FBS)

<br/>2. Dilute the recombinant protein in&nbsp; 0.9% saline solution or sterilize PBS to make a final concentration of 0.5 μg/μl

For 10 mice:&nbsp;<br/>Original protein concentration is 2 mg/ml, ''i.e.'', 2 μg/μl, take 125 μl, add 375 μl 0.9% saline or PBS solution. Mix. This is 0.5 μg/μl diluted antigen

Vortex Quickantibody adjuvant, add 550 μl Quickantibody adjuvant to the above 500 μl diluted antigen<br/>Final concentration: ~0.24 μg/μl, total 1050 μl.

3. Inject 100&nbsp; μl antigen intramuscularly in hind legs of each mouse, 50 μl to each leg.

It is normal to see precipitation in the antigen/adjuvant mixture. Mix well before drawing into a syringe and inject it as quick as possible after drawing into the syringe.



4. 3 weeks later, immunize mice again as above



5. 1 week after&nbsp;the 2nd immunization, draw blood from tails or eyes of mouse and test titer&nbsp;by ELISA,&nbsp;ELISA titers should be in the range of 1:1000~1:10000.

6. At week 6,&nbsp;immunize mice again as above.

7. Make hybridoma at week 7