Antibody Production using Titermax i. To blood sample Red top #15 scalpel Microtainer/Vacutainer™ Heat lamp in holder 22 Gauge needle 4x4 Restrainer Xylene/Oil of Wintergreen Folded Isopropyl Alcohol ii. To prepare antigen for injection Safety glasses 70% ethanol; cotton Gloves 22 Gauge needle(or smaller) to Titermax Gold™ Adjuvant inject I.M. / 18Gauge needle (or All plastic/luer lock syringes smaller) to inject S.Q. 22-16 needle to draw up Stainless connector sample Antigen NOTE: Titer-Max is available from CytRx Corporation, 150 Technology Parkway, Technology Park/Atlanta, Norcross, Georgia 30092 USA (phone 404-368-9500). iii. To prepare for cardiac exsanguination/euthanasia Ketamine/Rompun 1 cc/insulin syringes anesthetic mixture in 3cc 4x4 gauze swabs syringe Microtainer/Vacutainer™ holder, 22 Gauge needle and needles (20G x 1 ½ ) Fumehood and bell jar Pre-filled syringe of T-61 Towels Euthanasia Solution/Euthanyl Procedure Ascites Fluid Production and Collection The best mice to use for collection of ascites fluid are Balb/C females that are retired breeders (at least 6 months of age). These mice usually have less tumour formation and have greater abdominal muscle expansion for a larger volume of ascites. The mice are first primed with Freund’s Incomplete Adjuvant. This needs to be done at least 1 day or up to 3 days before injection with hybridoma cells. Primed mice can be used for up to one month. Mice in each cage do not have to be individually identified as the ascites fluid from each cage will be pooled. Injection of mice
Antibody Production using Titermax i. To blood sample Red top Microtainer/Vacutainer™ 22 Gauge needle Restrainer Folded #15 scalpel Heat lamp in holder 4x4 Xylene/Oil of Wintergreen Isopropyl Alcohol ii. To prepare antigen for injection Safety glasses Gloves Titermax Gold™ Adjuvant All plastic/luer lock syringes 22-16 needle to draw up sample 70% ethanol; cotton 22 Gauge needle(or smaller) to inject I.M. / 18Gauge needle (or smaller) to inject S.Q. Stainless connector Antigen NOTE: Titer-Max is available from CytRx Corporation, 150 Technology Parkway, Technology Park/Atlanta, Norcross, Georgia 30092 USA (phone 404-368-9500). iii. To prepare for cardiac exsanguination/euthanasia Ketamine/Rompun anesthetic mixture in 3cc syringe 22 Gauge needle Fumehood and bell jar Towels 1 cc/insulin syringes 4x4 gauze swabs Microtainer/Vacutainer™ holder, and needles (20G x 1 ½ ) Pre-filled syringe of T-61 Euthanasia Solution/Euthanyl Procedure Ascites Fluid Production and Collection The best mice to use for collection of ascites fluid are Balb/C females that are retired breeders (at least 6 months of age). These mice usually have less tumour formation and have greater abdominal muscle expansion for a larger volume of ascites. The mice are first primed with Freund’s Incomplete Adjuvant. This needs to be done at least 1 day or up to 3 days before injection with hybridoma cells. Primed mice can be used for up to one month. Mice in each cage do not have to be individually identified as the ascites fluid from each cage will be pooled. Injection of mice
B101(AntibodyProduction).docx