Mouse Anti-Chicken NCAM Hybridoma [E4]

Immunogen

Wheat germ agglutinin affinity purified chicken brain proteins immobilized on lectin beads

Fusion Species

Mouse X Mouse Hybridoma

Immunological Donor

BALB/c mouse spleen

Clone

E4

Cell Line Description

Cell line was established by fusion of mouse NS-1 cells with spleen cells from a BALB/c mouse immunized with wheat germ agglutinin affinity purified chicken brain proteins immobilized on lectin beads; described as secreting a mouse monoclonal antibody (igg) reacting with the cytoplasmic domain of the 140, 180 and 250 kda glycosylated neuronal cell surface protein N-CAM, preferentially in neurons of the chicken visual system like retina, tectum and other CNS regions and PNS; also reacting with mouse, goldfish and human N-CAM in paraformaldehyde-fixed cryosections and Western blots confirmed as murine with IEF of LDH, NP.
Cytogenetics: murine flat-moded hypertetraploid karyotype 98-10E4<4n> two centric fusion markers present.
Viruses: ELISA: reverse transcriptase positive; PCR: SMRV

Morphology

round, single cells growing in suspension and some slightly adherent

Growth Properties

suspension and some slightly adherent

Subculturing

Split ratio: 1: 5 to 1: 10, every 2-3 days; seed out at 0.5 x 10^6 cells/ml; Maintain at 0.5 x 10^6 cells/ml; Maximal density at1-2 x 10^6 cells/ml; Doubling time: 22 hours; Harvest: 0.5-1.0 x 10^6 cells/ml.
Some cells attach to the flask surface, but may be detached by tapping the flask.

Storage

Liquid nitrogen vapor phase.
Freezing medium: to complete growth medium(20%FBS), add 10%(v/v) DMSO