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From Peptide Antigens Provided by Customers To 2 Mouse Hybridomas (PE-CA)               

 Production Description
Step 1)Peptides conjugated to KLH for immunization and to BSA for ELISA
Step 2) Immunization of 6 Balb/c mice for each peptide-KLH conjugate
Step 3) ELISA monitoring of anti-sera development until titers reach >10,000
Step 4) Fusion of spleen cells from 1 mouse of highest ELISA titer with SP2/0
Step 5) 4-5 rounds of sub-cloning and ELISA screening to establish hybridomas
Step 6) Cryopreservation of cell lines
Step 7)Protein A/G affinity purification of ascites from 2 cell lines

 Order Today To Follow This Production Schedule
Date
Day
AbMART Track
Production Steps
01-06-2009
1
Order Date
 
01-13-2009
8
Peptide Receive

Receive peptides from customers;
Conjugation to KLH (for immunization) and to BSA (for ELISA);
6 mice ready (take pre-bleeds)
01-14-2009
9
Peptide Conjugation
01-15-2009
10
Adjuvant Addition
01-16-2009
11
Production Start
01-16-2009
11
IMM-1
1st Immunization (Complete Freud Adjuvant)
01-30-2009
25
IMM-2
2nd Immunization (Incomplete Freud Adjuvant)
02-13-2009
39
IMM-3
3rd Immunization (Incomplete Freud Adjuvant)
02-23-2009
49
BLD-M1
Bleed; ELISA
After 3rd Immunization, IF titers>10,000, go on to Fusion and hybridoma cloning: as early as
Date
02-28-2009
02-27-2009
53
IMM-4
4th Immunization (Incomplete Freud Adjuvant)
03-09-2009
63
BLD-M2
Bleed; ELISA
Day
54
03-13-2009
67
BST-F2
Final Boost (iv/ip)
03-16-2009
70
Fusion
 SP2/0 fusion with spleen cells from 1 mouse (plating in 10X 96-well plates)
03-26-2009
80
S1-ELISA 1-Sclone
S2-ELISA 2-Sclone
S3-ELISA 3-Sclone
S4-ELISA 4-Sclone
S5-ELISA
4-5 rounds of sub-cloning and ELISA screening
05-15-2009
130
05-29-2009
144
Cryopreservation
Expand cell lines and make frozen stocks
06-19-2009
165
Ascites Production/Purification
Protein A/G purification of ascites from 2 mice for each cell line
06-22-2009
168
Shipment
1) 2 hybridomas (Two 25 cm2 flasks with 2-5 x 106 cells in DMEM/high glucose/15% FBS);
2) 2-5 mg of Protein A/G purified mouse IgGs from ascites production;
3) ELISA data;
4) Cell line isotypes

2140 Xietu Road, Building 2 Shanghai, P.R. China 200032, Phone: (011)-86-21-64048593, Email: orders@ab-mart.com.cn