20F3 (α-GDGF2)
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HYBRIDOMA Volume 13, Number 3, 1994 Mary Ann Liebert, Inc., Publishers Monoclonal Antibodies 20F3 (a-GDGF2) ANTIGEN USED FOR IMMUNIZATION Partially purified glioma-derived growth factor-2 (GDGF2) from human malignant glioma cell line D-54MG. GDGF2 is mitogenic for BALB 3T3.A31 cells; high molecular weight (>I58 kD) gel-filtration fractions of preparative isoelectric focused culture supernates. (pi = 6.5) METHOD OF IMMUNIZATION Intramuscular (both thighs) injection of GDGF-2 emulsified in Ribi's MPL/TDM preparation; intraperitoneal injection of GDGF-2 in saline at 3-week intervals x3. Mice killed 4 days after last injection. PARENTAL CELL LINE USED FOR FUSION P3x63Ag8.653 SELECTION AND CLONING PROCEDURE Littlefield's HAT formulation; hybrids seeded in 20 96-well plates; colony supernates selected by screening for mouse Ig. Selected supernates screened by ELISA against immunogen separated by IEF or gel filtration and screened by ability to bind 35S-labeled glioma cell proteins. SPECIFICITY Binds to GDGF-2 but does not neutralize mitogenic activity directly. Can be used to immunoprecipitate GDGF-2 or to purify GDGF-2 by immuno affinity chromatography. Works poorly in Western blots of SDS-PAGE separated GDGF-2. SPECIFIC ANTIGEN IDENTIFIED GDGF-2 a high-molecular-weight (170 kD) mitogenic activity for BALB 3T3 cells; produced by human glioma cells. AVAILABILITY Tissue culture supernatant Ascitic fluid Hybridoma cells ÇYëT) No ÇYës^i No Yes CÑo~ ADDRESS REPRINT REQUESTS TO: G.Y. Gillespie, Ph.D. Division of Neurosurgery 65THT University of Alabama at Birmingham Birmingham, AL 35294-0006 (205) 934-7227; facsimile (205) 934-7676 HEAVY AND LIGHT CHAINS OF IMMUNOGLOBULIN IgG„k REFERENCES Lyon E and Gillespie GY: Hybridoma 1994;13:15-20. Lyon E and Gillespie GY: J Neuro-Oncol 1993;17:99-109. 252
Transcript of 20F3 (α-GDGF2)
HYBRIDOMAVolume 13, Number 3, 1994Mary Ann Liebert, Inc., Publishers
Monoclonal Antibodies
20F3 (a-GDGF2)
ANTIGEN USED FOR IMMUNIZATION
Partially purified glioma-derived growth factor-2 (GDGF2)from human malignant glioma cell line D-54MG. GDGF2 ismitogenic for BALB 3T3.A31 cells; high molecular weight(>I58 kD) gel-filtration fractions of preparative isoelectricfocused culture supernates. (pi = 6.5)
METHOD OF IMMUNIZATION
Intramuscular (both thighs) injection of GDGF-2 emulsified inRibi's MPL/TDM preparation; intraperitoneal injection ofGDGF-2 in saline at 3-week intervals x3. Mice killed 4 daysafter last injection.
PARENTAL CELL LINE USED FOR FUSION
P3x63Ag8.653
SELECTION AND CLONING PROCEDURE
Littlefield's HAT formulation; hybrids seeded in 20 96-wellplates; colony supernates selected by screening for mouse Ig.Selected supernates screened by ELISA against immunogenseparated by IEF or gel filtration and screened by ability to bind35S-labeled glioma cell proteins.
SPECIFICITY
Binds to GDGF-2 but does not neutralize mitogenic activitydirectly. Can be used to immunoprecipitate GDGF-2 or to purifyGDGF-2 by immuno affinity chromatography. Works poorly inWestern blots of SDS-PAGE separated GDGF-2.
SPECIFIC ANTIGEN IDENTIFIED
GDGF-2 a high-molecular-weight (170 kD) mitogenic activityfor BALB 3T3 cells; produced by human glioma cells.
AVAILABILITY
Tissue culture supernatantAscitic fluidHybridoma cells
ÇYëT) NoÇYës^i NoYes CÑo~
ADDRESS REPRINT REQUESTS TO:
G.Y. Gillespie, Ph.D.Division ofNeurosurgery65THTUniversity of Alabama at BirminghamBirmingham, AL 35294-0006(205) 934-7227;facsimile (205) 934-7676
HEAVY AND LIGHT CHAINS OFIMMUNOGLOBULIN
IgG„k
REFERENCES
Lyon E and Gillespie GY: Hybridoma 1994;13:15-20.Lyon E and Gillespie GY: J Neuro-Oncol 1993;17:99-109.
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