A procedure for the fusion of cells in suspension by means of polyethylene glycol

Hales, Anne

doi:10.1007/bf01551817

Cited by 66 publications

(19 citation statements)

References 5 publications

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“…A suspension of splenic lymphocytes was mixed with the rat myeloma line Y3.1.2.3.Ag (Galfre et al, 1979) and fused in polyethylene glycol (Hales, 1977). Hybrids were selected in HAT medium (Miller & Ruddle, 1976) and cloned in agar.…”

mentioning

confidence: 99%

Localisation of metastatic carcinoma by a radiolabelled monoclonal antibody

Smedley¹,

Finan²,

Lennox³

et al. 1983

Br J Cancer

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mentioning

confidence: 99%

Localisation of metastatic carcinoma by a radiolabelled monoclonal antibody

Smedley¹,

Finan²,

Lennox³

et al. 1983

Br J Cancer

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“…Before extraction of DNA from the hybrid cells, we confirmed the presence of a single mouse X chromosome in the background of Chinese hamster chromosomes by means of an R-banding method involving bromodeoxyuridine labeling and acridine orange staining. S194 cells were fused with splenocytes from a young adult mouse with T(X;4)37H (hereafetr shortened to T37H) translocation, a generous gift of Dr. A. G. Searle (MRC Radiobiology Unit, Chilton, UK), by the method described by Hales (9). A number of HAT-resistant clones were isolated by limiting dilution and one of the clones was designated as MHH-1.…”

Section: Cell Culture and Cell Fusionmentioning

confidence: 99%

Construction of a DNA Library Enriched with Mouse 4^xChromosome of T(X;4)37H Translocation

et al. 1990

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Normal mouse chromosomes are routinely separated into only 5 peaks by the current flow cytometry. Since this limited resolution hindered isolation of the normal mouse X chromosome with an appropriate purity, we attempted to sort the mouse 4x chromosome, a larger translocation chromosome of T(X;4)37H, consisting of nearly the entire chromosome 4 and chromosome X by flow cytometry. To obtain a large number of cells having 4x chromosome for flow sorting, we isolated a somatic hybrid cell line MHH‐1 formed between S194 myelome cell line and normal splenocytes from a male mouse carrying T(X;4)37H. Flow karyotyping of propidium iodide‐stained chromosomes from MHH‐1 cell line revealed an additional peak containing 4x chromosomes at about 80%. DNA purified from sorted 4x chromosomes was cloned into phage lambda gtWES after complete digestion with EcoRl restriction endonuclease. Thus a 4x chromosome‐enriched library of about 4.4 × 104 recombinant phages was made and 13 single copy DNA clones specific to the X chromosome were isolated from the library so far.

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“…lo7 myeloma cells (P3-NS-1-Ag 4-1) [I71 and 3 . lo8 spleen cells from the hyperimmunized mouse were fused with the aid of poly(ethyleneglyco1) (PEG 1000, Baker, Deventer, The Netherlands) as described in [18]. After one day the medium was replaced by one containing hypoxanthineaminopterin-thymidine (HAT medium [I 91) and the cell suspension was plated at a density of 4 .…”

Section: Cell Culture Hybridization and Cloningmentioning

confidence: 99%

Monoclonal Antibodies against Human β‐Glucocerebrosidase

Barneveld

Tegelaers

Ginns

et al. 1983

European Journal of Biochemistry

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Monoclonal antibodies were obtained against the membrane‐bound lysosomal enzyme β‐glucocerebrosidase (acid β‐glucosidase), which is deficient in Gaucher's disease. BALB/c mice were immunized with homogeneous enzyme protein extracted from a sodium dodecyl sulphate/polyacrylamide gel. The mice were subsequently hyperimmunized with partially purified enzyme prior to fusion of spleen cells with myeloma cells. After fusion, 32 primary hybrid cell populations were obtained which continued to produce antibodies against β‐glucocerebrosidase after prolonged time of culture. All antibodies reacted with both native and denatured enzyme. Four primary cell populations were subcloned and the antibodies produced were characterized. The antibodies were all four of the IgG1 subclass. Three of these antibodies bind to protein A whereas one does not. The results of binding assays indicated that three of the antibodies react with the same antigenic domain (epitope 1), but the fourth with a different one (epitope 2). Probably two antigenic determinants are present in epitope 1 since one of the antibodies with specificity for epitope 1 is inactivated after iodination by the chloramine‐T procedure whereas a second one is not.

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A procedure for the fusion of cells in suspension by means of polyethylene glycol

Cited by 66 publications

References 5 publications

Localisation of metastatic carcinoma by a radiolabelled monoclonal antibody

Localisation of metastatic carcinoma by a radiolabelled monoclonal antibody

Construction of a DNA Library Enriched with Mouse 4^xChromosome of T(X;4)37H Translocation

Monoclonal Antibodies against Human β‐Glucocerebrosidase

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A procedure for the fusion of cells in suspension by means of polyethylene glycol

Cited by 66 publications

References 5 publications

Localisation of metastatic carcinoma by a radiolabelled monoclonal antibody

Localisation of metastatic carcinoma by a radiolabelled monoclonal antibody

Construction of a DNA Library Enriched with Mouse 4xChromosome of T(X;4)37H Translocation

Monoclonal Antibodies against Human β‐Glucocerebrosidase

Contact Info

Product

Resources

About

Construction of a DNA Library Enriched with Mouse 4^xChromosome of T(X;4)37H Translocation