Editing Hybridoma production (section)

=== Materials ===

{| border="1"
|-
| '''Name'''
| '''Company'''
| '''Catalog Number'''
| '''Comments'''
|-
| glutaraldehyde
| Sigma Aldrich
| G5882
|-
| ''N''-(3-dimethylaminopropyl)-''N''-ethylcarbodiimide (EDC)
| Sigma Aldrich
| 39391-10ML
|-
| Sulfo-GMBS
| Perbio Science Germany
| 22324
|-
| ovalbumin
| Sigma Aldrich
| A5503
|-
| bovine serum albumin
| Sigma Aldrich
| A2153
|-
| keyhole limpet hemocyanin
| Sigma Aldrich
| H8283
|-
| Falcon tubes 15 mL
| Biochrom GmbH
| P91015
|-
| reaction vials, 1.5 mL
| Carl Roth GmbH & C0.KG
| CNT2.1
|-
| hollow needle
| Carl Roth GmbH & C0.KG
| C724.1
|-
| glass wool
| Carl Roth GmbH & C0.KG
| 6574.1
|-
| Sephadex G25 coarse
| Sigma Aldrich
| GE-17-0034-02
|-
| Freund´s adjuvant, complete
| Sigma Aldrich
| F5881-10ML
|-
| ELISA plates, 96 well
| Greiner bio-one
| 655101
|-
| neonatal calf serum
| Biochrom GmbH
| S1025
|-
| TipOne Tips 1,000 µL
| Starlab
| S1111-2021
|-
| Pipette tips 200 µL
| Greiner bio-one
| 739291
|-
| HRP-conjugated goat-anti-mouse IgG antibody
| Dianova
| 115-035-003
|-
| tetramethylbenzidine
| Carl Roth GmbH & C0.KG
| 6350.2
|-
| Natriumdihydrogenphosphat
| Carl Roth GmbH & C0.KG
| K300.2
|-
| peroxide/urea
|-
| sulphuric acid
| Carl Roth GmbH & C0.KG
| 4623.3
|-
| RPMI 1640
| Life technologies GmbH
| 31870074
|-
| L-glutamine
| Carl Roth GmbH & C0.KG
| HN08.2
|-
| beta-mercaptoethanol
| Sigma Aldrich
| M6250
|-
| fetal calf serum
| Invitrogen
| 10270106
|-
| TC-flask 25 cm<sup>2</sup>
| Peske GmbH
| 86-V025
|-
| TC-flask 75 cm<sup>2</sup>
| Peske GmbH
| 86-V075
|-
| ethanol, 96%
| Carl Roth GmbH & C0.KG
| P075.1
|-
| cell strainer
| VWR international
| 734-0002
|-
| Falcon tubes 50 mL
| Biochrom GmbH
| P91050
|-
| PEG 8000
| Sigma Aldrich
| 1546605
|-
| electroporation cuvette, 2 mm
| Biodeal Handelsvertretung Edelmann e.K.
| EKL2,25
|-
| hypoxanthine
| Sigma Aldrich
| H9636-25G
|-
| azaserine
| Sigma Aldrich
| A4142
|-
| thymidine
| USB Europa GmbH
| 22305 1 GM
|-
| TC-plates 96 well
| Biochrom GmbH
| P92696
|-
| TC-plates 24 well
| Biochrom GmbH
| P92424
|-
| cryotubes, 1 mL
| Sigma Aldrich
| V7384-1CS
|-
| dimethylsulfoxide
| Carl Roth GmbH & C0.KG
| 4720.1
|-
| protein A sepharose
| Sigma Aldrich
| P3391-1G
|-
| SDS sample loading buffer, Roti-Load 1
| Carl Roth GmbH & C0.KG
| K929.1
|-
| unstained protein ladder
| BioRad Laboratories
| 161-0363
|-
| comassie brilliant blue R-250
| BioRad Laboratories
| 161-0406
|}

References

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#Hanack, K., Messerschmidt, K., Listek, M. Antibodies and Selection. ''Protein Targeting Compounds''. Böldicke, T. Springer Verlag. Berlin Heidelberg. 11-22 (2015).
#Walsh, G. Biopharmaceutical benchmarks. ''Nat Biotechnol''. '''32''', (10), 992-1000 (2014).
#Pauly, D., Hanack, K. How to avoid pitfalls in antibody use. ''F1000 Res''. '''7''', (4), 691-698 (2015).
#Bradbury, A., Plückthun, A. Reproducibility: Standardize antibodies used in research. ''Nature''. '''5''', (518), 27-29 (2015).
#Polakiewicz, R. D. Antibodies: The solution is validation. ''Nature''. '''26''', (518), 483 (2015).
#Baker, M. Antibody anarchy: A call to order. ''Nature''. '''26''', (527), 545-551 (2015).
#Smith, G. P. Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. ''Science''. '''228''', (4705), 1315-1317 (1985).
#Kearney, J. F., Radbruch, A., Liesegang, B., Rajewsky, K. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. ''J Immunol''. '''123''', (4), 1548-1550 (1979).
#Shulman, M., Wilde, C. D., Köhler, G. A better cell line for making hybridomas secreting specific antibodies. ''Nature''. '''16''', (276), 269-270 (1978).
#Stoicheva, N. G., Hui, S. W. Electrically induced fusion of mammalian cells in the presence of polyethylene glycol. ''J Membr Biol''. '''141''', (2), 177-182 (1994).
#Osterhaus, A. D., Uytdehaag, A. G. Current developments in hybridoma technology. ''Tijdschr Diergeneeskd''. '''15''', (110), 835-839 (1985).
#Migneault, I., Dartiguenave, C., Bertrand, M. J., Waldron, K. C. Glutaraldehyde: behavior in aqueous solution, reaction with proteins, and application to enzyme crosslinking. ''Biotechniques''. '''37''', (5), 798-802 (2004).
#Vitetta, E. S., Baur, S., Uhr, J. W. Cell Surface Immunoglobulin II. Isolation and characterization of immunoglobulin from mouse splenic lymphocytes. ''Journal of experimental medicine''. '''134''', 242-264 (1971).
#Laemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. ''Nature''. '''227''', (5259), 680-685 (1970).
#Browne, S. M., Al-Rubeai, M. Selection methods for high-producing mammalian cell lines. ''Trends Biotechnol''. '''25''', (9), 425-432 (2007).
#Holmes, P., Al-Rubeai, M. Improved cell line development by a high throughput affinity capture surface display technique to select for high secretors. ''J Immunol Methods''. '''19''', (230), 141-147 (1999).
#Weaver, J. C., McGrath, P., Adams, S. Gel microdrop technology for rapid isolation of rare and high producer cells. ''Nat Med''. '''3''', (5), 583-585 (1997).
#Messerschmidt, K., Hempel, S., Holzlöhner, P., Ulrich, R. G., Wagner, D., Heilmann, K. IgA antibody production by intrarectal immunization of mice using recombinant major capsid protein of hamster polyomavirus. ''Eur J Microbiol Immunol''. '''2''', (3), 231-238 (2012).
#Listek, M., Micheel, B., Hanack, K. Biomoleküle freisetzende zelle und deren selektion mittels eines oberflächenproteins. ''neweramabs GmbH''. PCT/DE2014/000205 (2014).
[[Category:Pages with broken file links]] [[Category:Antibodies]] [[Category:Protocols]]