Since collagen has an unique amino acid composition which is rich in Glycine and Proline, general protein assays do not show correct collagen levels when…
Month: January 2021
Hybridoma Cloning
After cell fusion, when we have positive wells, the next step is cloning cells. Sometimes, the positive wells have several colonies in the wells. Generally,…
Making emulsion – a syringe method
Generally, emulsion can be prepared using a two-syringe method. Two syringes are connected with a connector. Each syringe contains equal volume of antigen solution and…
Purification of antibodies
Antibodies, especially monoclonal antibodies, can be purified by Protein A, Protein G, or Protein L which binds to IgG specifically. However, when we need to…
Denatured collagen analysis
Native collagen can be evaluated using immunoassays with monoclonal antibodies (mAbs) against collagen. Most mAbs were developed by native collagen immunization. Denatured collagen is easy…
Low OD values on ELISA 1
Enzyme-linked solvent assay (ELISA) is a very useful and popular method to analyze biological analytes. However, we sometimes have problems with not paying attention each…
Conditioning hybridoma culture
Culturing hybridoma is sometimes challenging. The growth rate and antibody production vary in individual clones. We generally choose hybridoma who has fast growth and high…
Rat Collagen-induced Arthritis (CIA)
Rat CIA can be induced by immunization of type II collagen (Porcine or Bovine) emulsified with Incomplete adjuvant (IFA). The injection can be SC injection…
Collagen solubilization
Collagen sample preparation (1) Acid soluble: Works only for skin samples from young animals. Neutral buffer soluble: Works partially for samples after acid or pepsin…
Collagen induced arthritis in B6 mice
Since C57BL/6 mice are a low susceptible strain to collagen induced arthritis (CIA), the arthritis model requires high concentration of M.tuberculosis in Complete Freund’s Adjuvant…
Oral sample preparation
Oral samples can be used for cytokine analysis. However, sample preparation must be paid attention. Gingival crevicular fluid (GCF) The gingival crevicular fluid is collected…