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From cDNA Clones Provided by Customers To 2 Mouse Hybridomas (CD-CL)               

 Production Description
Step 1) Build expression plasmid based on cDNA clones provided by customers
Step 2) Express and purify tagged (His- or GST-) recombinant proteins
Step 3) Immunization of 6 Balb/c mice for each peptide-KLH conjugate
Step 4) ELISA monitoring of anti-sera development until titers reach >10,000
Step 5) Fusion of spleen cells from 1 mouse of highest ELISA titer with SP2/0
Step 6) 4-5 rounds of sub-cloning and ELISA screening to establish hybridomas
Step 7) Cryopreservation of cell lines

 Order Today To Follow This Production Schedule
Date
Day
AbMART Track
Production Steps
11-21-2008
1
Order Date
 
11-24-2008
4
Peptide Design
Design gene cloning strategy for expression plasmid vector construction;
recombinant protein production and purification;
6 mice ready (Pre-bleed is done)
12-29-2008
39
Protein Production
01-01-2009
42
Protein Receive
01-02-2009
43
Adjuvant Addition
01-03-2009
44
Production Start
01-03-2009
44
IMM-1
1st Immunization (Complete Freud Adjuvant)
01-17-2009
58
IMM-2
2nd Immunization (Incomplete Freud Adjuvant)
01-31-2009
72
IMM-3
3rd Immunization (Incomplete Freud Adjuvant)
02-10-2009
82
BLD-M1
Bleed; ELISA
After 3rd Immunization, IF titers>10,000, go on to Fusion and hybridoma cloning: as early as
Date
02-15-2009
02-14-2009
86
IMM-4
4th Immunization (Incomplete Freud Adjuvant)
02-24-2009
96
BLD-M2
Bleed; ELISA
Day
87
02-28-2009
100
BST-F2
Final Boost (iv/ip)
03-03-2009
103
Fusion
SP2/0 fusion with spleen cells from 1 mouse (plating in 10X 96-well plates)
03-13-2009
113
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
05-02-2009
163
05-16-2009
177
Cryopreservation
Expand cell lines and make frozen stocks
05-21-2009
182
Shipment
1) 2 hybridomas (Two 25 cm2 flasks with 2-5 x 106 cells in DMEM/high glucose/15% FBS);
2) ELISA data;
3) Cell line isotypes
4) 1-3 mg of recombinant protein

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