mouse anti rat lambda light chain hrp  (Bio-Rad)

Journal: Current Issues in Molecular Biology

Article Title: Development of a Novel Anti-CD44 Variant 4 Monoclonal Antibody C 44 Mab-108 for Immunohistochemistry

doi: 10.3390/cimb45030121

Figure Lengend Snippet: A schematic procedure of anti-CD44v4 mAbs production. ( A ) The mice were intraperitoneally immunized with the CD44v4 peptide conjugated with keyhole limpet hemocyanin (KLH). ( B ) The spleen cells were fused with P3U1 cells, and hybridomas were produced. ( C ) The supernatants were subsequently screened using enzyme-linked immunosorbent assay (ELISA) with the CD44v4 peptide, followed by flow cytometry using CHO/CD44v3-10 and CHO-K1 cells. CHO/CD44v3-10-reactive and parental CHO-K1-non-reactive supernatants were determined to be positive for CD44v3-10. ( D ) To establish single clones, limiting dilution was performed. A clone C 44 Mab-108 (IgG 1 , kappa) was finally established.

Article Snippet: After blocking with 1% BSA in PBST for 30 min at 37 °C, the plates were incubated with the supernatant of C 44 Mab-108, followed by incubation with peroxidase-conjugated anti-mouse immunoglobulins, anti-mouse heavy chains (IgG 1 , IgG 2a , IgG 2b , IgG 3 , and IgM; SouthernBiotech, Birmingham, AL, USA), or anti-mouse light chains (kappa and lambda; SouthernBiotech).

Techniques: Produced, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Clone Assay