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01000caa a22002652 4500 |
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NLM097594199 |
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DE-627 |
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20250131221031.0 |
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tu |
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231222s1998 xx ||||| 00| ||eng c |
| 028 |
5 |
2 |
|a pubmed25n0326.xml
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|a (DE-627)NLM097594199
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|a (NLM)9821584
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|a DE-627
|b ger
|c DE-627
|e rakwb
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| 041 |
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|a eng
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| 100 |
1 |
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|a O'Reilly, L A
|e verfasserin
|4 aut
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| 245 |
1 |
0 |
|a Rapid hybridoma screening method for the identification of monoclonal antibodies to low-abundance cytoplasmic proteins
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| 264 |
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1 |
|c 1998
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| 336 |
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|a Text
|b txt
|2 rdacontent
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| 337 |
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|a ohne Hilfsmittel zu benutzen
|b n
|2 rdamedia
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| 338 |
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|a Band
|b nc
|2 rdacarrier
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| 500 |
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|a Date Completed 19.01.1999
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| 500 |
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|a Date Revised 28.09.2018
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| 500 |
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|a published: Print
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| 500 |
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|a Citation Status MEDLINE
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| 520 |
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|a Screening assays are the most time-consuming and labor-intensive part of generating monoclonal antibodies (MAbs). Antibodies identified by enzyme-linked immunosorbent assay (ELISA) screening often are not suitable for their intended application such as immunofluorescence staining. We describe here a rapid and efficient flow cytometric screening procedure for the identification of MAbs directed against low-abundance cytoplasmic proteins, in our case, the pro-apoptotic molecule Bim. Cells from an equal mixture of a parental cell line and a subline expressing Bim were fixed, permeabilized and incubated with hybridoma supernatants. The supernatants were derived from a fusion of Sp2/0 plasmacytoma cells and spleen cells from a rat immunized with recombinant glutathione-S-transferase (GST)-BimL fusion protein. Secondary staining with fluorochrome-labeled anti-rat Ig antibodies allowed detection of clones expressing Bim-specific antibodies. The screening procedure was rapid and efficient, and most monoclonal antibodies identified were proven to be useful for immunofluorescence staining and other applications
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| 650 |
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4 |
|a Research Support, Non-U.S. Gov't
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| 650 |
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4 |
|a Technical Report
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| 650 |
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4 |
|a Journal Article
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| 650 |
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7 |
|a Antibodies, Monoclonal
|2 NLM
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| 650 |
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7 |
|a Apoptosis Regulatory Proteins
|2 NLM
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| 650 |
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7 |
|a BCL2L11 protein, human
|2 NLM
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| 650 |
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7 |
|a Bcl-2-Like Protein 11
|2 NLM
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| 650 |
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7 |
|a Bcl2l11 protein, mouse
|2 NLM
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| 650 |
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7 |
|a Bcl2l11 protein, rat
|2 NLM
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| 650 |
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7 |
|a Carrier Proteins
|2 NLM
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| 650 |
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7 |
|a Membrane Proteins
|2 NLM
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| 650 |
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7 |
|a Proto-Oncogene Proteins
|2 NLM
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| 650 |
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7 |
|a Recombinant Fusion Proteins
|2 NLM
|
| 700 |
1 |
|
|a Cullen, L
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Moriishi, K
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a O'Connor, L
|e verfasserin
|4 aut
|
| 700 |
1 |
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|a Huang, D C
|e verfasserin
|4 aut
|
| 700 |
1 |
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|a Strasser, A
|e verfasserin
|4 aut
|
| 773 |
0 |
8 |
|i Enthalten in
|t BioTechniques
|d 1993
|g 25(1998), 5 vom: 15. Nov., Seite 824-30
|w (DE-627)NLM012627046
|x 0736-6205
|7 nnns
|
| 773 |
1 |
8 |
|g volume:25
|g year:1998
|g number:5
|g day:15
|g month:11
|g pages:824-30
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|d 25
|j 1998
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|c 11
|h 824-30
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