3G8- pronephros and otic vesicle - Revision history

136.159.234.129 at 20:24, 28 September 2011

2011-09-28T20:24:15Z

← Older revision		Revision as of 20:24, 28 September 2011
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	'''Xenbase Gene Page:'''		'''Xenbase Gene Page:'''

	'''Epitope:'''		'''Epitope:''' Unknown- but localized to apical surface/brush border.

	'''Host Species/Isotype:''' mouse monoclonal IgG		'''Host Species/Isotype:''' mouse monoclonal IgG

68.147.37.142 at 17:40, 20 July 2011

2011-07-20T17:40:40Z

← Older revision		Revision as of 17:40, 20 July 2011
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	'''Antigen Species:''' Xenopus laevis		'''Antigen Species:''' Xenopus laevis

	'''Applications:''' IF		'''Applications:''' Immunofluorescence, immunohistochemistry

	'''Antibody Name/Catalog Number:''' 3G8		'''Antibody Name/Catalog Number:''' 3G8

68.147.37.142 at 17:39, 20 July 2011

2011-07-20T17:39:24Z

← Older revision		Revision as of 17:39, 20 July 2011
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	'''Antigen:''' dissected ~~pro nephroi~~, specific target unknown		'''Antigen:''' dissected pronephroi, specific target unknown

	'''Xenbase Gene Page:'''		'''Xenbase Gene Page:'''

imported>Pvize at 22:32, 8 March 2011

2011-03-08T22:32:11Z

← Older revision		Revision as of 22:32, 8 March 2011
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	'''Host Species/Isotype:''' mouse monoclonal IgG		'''Host Species/Isotype:''' mouse monoclonal IgG

	'''Antigen Species:'''		'''Antigen Species:''' Xenopus laevis

	'''Applications:''' IF		'''Applications:''' IF

imported>Pvize at 22:31, 8 March 2011

2011-03-08T22:31:41Z

← Older revision		Revision as of 22:31, 8 March 2011
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	'''Antibody Name/Catalog Number:''' 3G8		'''Antibody Name/Catalog Number:''' 3G8

	'''Supplier:''' ~~Liz Jones (ea@dna~~.~~bio.warwick~~.ac.uk~~) , 10 ml 3x. Sent on dry ice so please provide account number for shipping costs.~~		'''Supplier:''' EXRC [http://www.port.ac.uk/research/exrc/antibodies/Antibodies/]

	'''Comments:''' IP unknown, does not work on Westerns		'''Comments:''' IP unknown, does not work on Westerns

			[[File:3G8.jpg]]
			Note- the blue stain in this image is by antibody 3G8, the red stain is by antibody 4A6 [http://www.xenbase.org/xenwiki/index.php/Monoclonal_4A6_-_pronephros]

	'''Contributor:''' Generated and characterized by Elizabeth Jones [http://www.xenbase.org:80/community/person.do?method=display&personId=753]. References: Dev Bio 171:531- [http://www.xenbase.org/literature/article.do?method=display&articleId=19219]		'''Contributor:''' Generated and characterized by Elizabeth Jones [http://www.xenbase.org:80/community/person.do?method=display&personId=753]. References: Dev Bio 171:531- [http://www.xenbase.org/literature/article.do?method=display&articleId=19219]

205.142.197.33 at 19:01, 8 March 2011

2011-03-08T19:01:24Z

← Older revision		Revision as of 19:01, 8 March 2011
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	'''Antigen:''' dissected pro nephroi, specific target unknown		'''Antigen:''' dissected pro nephroi, specific target unknown

			'''Xenbase Gene Page:'''

	'''Epitope:'''		'''Epitope:'''
Line 9:		Line 11:
	'''Applications:''' IF		'''Applications:''' IF

	'''Antibody Name:''' 3G8		'''Antibody Name/Catalog Number:''' 3G8

	'''Supplier:''' Liz Jones (ea@dna.bio.warwick.ac.uk) , 10 ml 3x. Sent on dry ice so please provide account number for shipping costs.		'''Supplier:''' Liz Jones (ea@dna.bio.warwick.ac.uk) , 10 ml 3x. Sent on dry ice so please provide account number for shipping costs.
Line 17:		Line 19:
	'''Contributor:''' Generated and characterized by Elizabeth Jones [http://www.xenbase.org:80/community/person.do?method=display&personId=753]. References: Dev Bio 171:531- [http://www.xenbase.org/literature/article.do?method=display&articleId=19219]		'''Contributor:''' Generated and characterized by Elizabeth Jones [http://www.xenbase.org:80/community/person.do?method=display&personId=753]. References: Dev Bio 171:531- [http://www.xenbase.org/literature/article.do?method=display&articleId=19219]

	'''Additional Information:'''		'''Additional Information:''' '''Developmental Window:''' First detects target at stage 31, continues until latest stages tested. As this antibody stained only the lumen of the pronephric proximal tubules the shape of the organ looks odd in early stages and only begins to look like a pronephros at around stage 34 when a well luminized epithelial tubule network has formed.

	'''Developmental Window:''' First detects target at stage 31, continues until latest stages tested. As this antibody stained only the lumen of the pronephric proximal tubules the shape of the organ looks odd in early stages and only begins to look like a pronephros at around stage 34 when a well luminized epithelial tubule network has formed.

	'''Stains:''' proximal pronephric nephron, nephrostomes and otic vesicle,		'''Stains:''' proximal pronephric nephron, nephrostomes and otic vesicle,

	'''Fixatives:''' formalin, MEMFA, TCA are all OK. Following fixation sample must be dehydrated with methanol for the antibody to work optimally. Others not tested. The target degrades over time in alcohol, so samples should be stored for less than one month post-fixation.		'''Fixatives:''' formalin, MEMFA, TCA are all OK. Following fixation sample must be dehydrated with methanol for the antibody to work optimally. Others not tested. The target degrades over time in alcohol, so samples should be stored for less than one month post-fixation.

	'''Sensitive to bleaching:''' No, though store time in methanol may be shortened by bleaching		'''Sensitive to bleaching:''' No, though store time in methanol may be shortened by bleaching

	'''Functions post in situ?:''' No		'''Functions post in situ?:''' No

205.142.197.33 at 13:49, 8 March 2011

2011-03-08T13:49:57Z

← Older revision		Revision as of 13:49, 8 March 2011
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	'''Antigen:''' dissected pro nephroi, specific target unknown		'''Antigen:''' dissected pro nephroi, specific target unknown

	'''Epitope:'''		'''Epitope:'''

	'''Host Species/Isotype:''' mouse monoclonal IgG		'''Host Species/Isotype:''' mouse monoclonal IgG

	'''Antigen Species:'''		'''Antigen Species:'''

	'''Applications:''' IF		'''Applications:''' IF

	'''Antibody Name:''' 3G8		'''Antibody Name:''' 3G8

	'''Supplier:''' Liz Jones (ea@dna.bio.warwick.ac.uk) , 10 ml 3x. Sent on dry ice so please provide account number for shipping costs.		'''Supplier:''' Liz Jones (ea@dna.bio.warwick.ac.uk) , 10 ml 3x. Sent on dry ice so please provide account number for shipping costs.

	'''Comments:''' IP unknown, does not work on Westerns		'''Comments:''' IP unknown, does not work on Westerns

	'''Contributor:''' Generated and characterized by Elizabeth Jones [http://www.xenbase.org:80/community/person.do?method=display&personId=753]. References: Dev Bio 171:531- [http://www.xenbase.org/literature/article.do?method=display&articleId=19219]		'''Contributor:''' Generated and characterized by Elizabeth Jones [http://www.xenbase.org:80/community/person.do?method=display&personId=753]. References: Dev Bio 171:531- [http://www.xenbase.org/literature/article.do?method=display&articleId=19219]

			'''Additional Information:'''
	~~applications abbreviations~~:
	~~WB - Western blotting,~~		'''Developmental Window:''' First detects target at stage 31, continues until latest stages tested. As this antibody stained only the lumen of the pronephric proximal tubules the shape of the organ looks odd in early stages and only begins to look like a pronephros at around stage 34 when a well luminized epithelial tubule network has formed.
	~~IP - immunoprecipitation,~~
	~~IF - immunofluorescence,~~		'''Stains:''' proximal pronephric nephron, nephrostomes and otic vesicle,
	~~IHC - immunohistochemistry,~~
	~~EMSA - electromobility shift assay,~~		'''Fixatives:''' formalin, MEMFA, TCA are all OK. Following fixation sample must be dehydrated with methanol for the antibody to work optimally. Others not tested. The target degrades over time in alcohol, so samples should be stored for less than one month post-fixation.
	~~ChIP - chromatin immunoprecipitation,~~
	~~FCM - flow cytometry~~		'''Sensitive to bleaching:''' No, though store time in methanol may be shortened by bleaching

			'''Functions post in situ?:''' No
	~~Additional Comments:~~
			'''Optimized conditions:''' Very clean antibody, very stable, can be reused. Development in BCIP/NBT usually takes 30 minutes, in red, about 60 minutes. Works well in immunofluorescense. As the pronephros is only covered by a thin transparent epidermis it is not necessary to clear stained embryos. In cleared embryos both pronephroi are often in view, one in focus and one blurred, and this can make photography difficult see for example stage 32 sample above.
	'''Developmental Window:''' First detects target at stage 31, continues until latest stages tested. As this antibody stained only the lumen of the pronephric proximal tubules the shape of the organ looks odd in early stages and only begins to look like a pronephros at around stage 34 when a well luminized epithelial tubule network has formed.
			'''Other species:''' works on other anurans (Bombina), and also on axolotls when formalin plus 10% DMSO is used as the fixative, but it isn't nearly as clean as with Xenopus or Bombina. This is probably just a protocol problem. Reported to work on zebrafish.
	'''Stains:''' proximal pronephric nephron, nephrostomes and otic vesicle,
			'''XAO:'''
	'''Fixatives:''' formalin, MEMFA, TCA are all OK. Following fixation sample must be dehydrated with methanol for the antibody to work optimally. Others not tested. The target degrades over time in alcohol, so samples should be stored for less than one month post-fixation.
			'''Reference(s):''' Dev Bio 171:531- [http://www.xenbase.org/literature/article.do?method=display&articleId=19219]
	'''Sensitive to bleaching:''' No, though store time in methanol may be shortened by bleaching
			applications abbreviations:
	'''Functions post in situ?:''' No
			WB - Western blotting,
	'''Optimized conditions:''' Very clean antibody, very stable, can be reused. Development in BCIP/NBT usually takes 30 minutes, in red, about 60 minutes. Works well in immunofluorescense. As the pronephros is only covered by a thin transparent epidermis it is not necessary to clear stained embryos. In cleared embryos both pronephroi are often in view, one in focus and one blurred, and this can make photography difficult see for example stage 32 sample above.
			IP - immunoprecipitation,
	'''Other species:''' works on other anurans (Bombina), and also on axolotls when formalin plus 10% DMSO is used as the fixative, but it isn't nearly as clean as with Xenopus or Bombina. This is probably just a protocol problem. Reported to work on zebrafish.
			IF - immunofluorescence,
	'''XAO:'''
			IHC - immunohistochemistry,
	~~References~~: Dev Bio 171:531- [http://www.xenbase.org/literature/article.do?method=display&articleId=19219]
			EMSA - electromobility shift assay,

			ChIP - chromatin immunoprecipitation,

			FCM - flow cytometry

205.142.197.91 at 19:28, 4 February 2011

2011-02-04T19:28:45Z

← Older revision		Revision as of 19:28, 4 February 2011
Line 27:		Line 27:
	FCM - flow cytometry		FCM - flow cytometry

	Additional Comments:
			Additional Comments:

	'''Developmental Window:''' First detects target at stage 31, continues until latest stages tested. As this antibody stained only the lumen of the pronephric proximal tubules the shape of the organ looks odd in early stages and only begins to look like a pronephros at around stage 34 when a well luminized epithelial tubule network has formed.		'''Developmental Window:''' First detects target at stage 31, continues until latest stages tested. As this antibody stained only the lumen of the pronephric proximal tubules the shape of the organ looks odd in early stages and only begins to look like a pronephros at around stage 34 when a well luminized epithelial tubule network has formed.

205.142.197.91: Created page with "'''Antigen:''' dissected pro nephroi, specific target unknown '''Epitope:''' '''Host Species/Isotype:''' mouse monoclonal IgG '''Antigen Species:''' '''..."

2011-02-04T19:27:04Z

Created page with "'''Antigen:''' dissected pro nephroi, specific target unknown '''Epitope:''' '''Host Species/Isotype:''' mouse monoclonal IgG '''Antigen Species:''' '''..."

New page

'''Antigen:''' dissected pro nephroi, specific target unknown

'''Epitope:'''

'''Host Species/Isotype:''' mouse monoclonal IgG

'''Antigen Species:'''

'''Applications:''' IF

'''Antibody Name:''' 3G8

'''Supplier:''' Liz Jones (ea@dna.bio.warwick.ac.uk) , 10 ml 3x. Sent on dry ice so please provide account number for shipping costs.

'''Comments:''' IP unknown, does not work on Westerns

'''Contributor:''' Generated and characterized by Elizabeth Jones [http://www.xenbase.org:80/community/person.do?method=display&personId=753]. References: Dev Bio 171:531- [http://www.xenbase.org/literature/article.do?method=display&articleId=19219]

applications abbreviations:
WB - Western blotting,
IP - immunoprecipitation,
IF - immunofluorescence,
IHC - immunohistochemistry,
EMSA - electromobility shift assay,
ChIP - chromatin immunoprecipitation,
FCM - flow cytometry

Additional Comments:
'''Developmental Window:''' First detects target at stage 31, continues until latest stages tested. As this antibody stained only the lumen of the pronephric proximal tubules the shape of the organ looks odd in early stages and only begins to look like a pronephros at around stage 34 when a well luminized epithelial tubule network has formed.

'''Stains:''' proximal pronephric nephron, nephrostomes and otic vesicle,

'''Fixatives:''' formalin, MEMFA, TCA are all OK. Following fixation sample must be dehydrated with methanol for the antibody to work optimally. Others not tested. The target degrades over time in alcohol, so samples should be stored for less than one month post-fixation.

'''Sensitive to bleaching:''' No, though store time in methanol may be shortened by bleaching

'''Functions post in situ?:''' No

'''Optimized conditions:''' Very clean antibody, very stable, can be reused. Development in BCIP/NBT usually takes 30 minutes, in red, about 60 minutes. Works well in immunofluorescense. As the pronephros is only covered by a thin transparent epidermis it is not necessary to clear stained embryos. In cleared embryos both pronephroi are often in view, one in focus and one blurred, and this can make photography difficult see for example stage 32 sample above.

'''Other species:''' works on other anurans (Bombina), and also on axolotls when formalin plus 10% DMSO is used as the fixative, but it isn't nearly as clean as with Xenopus or Bombina. This is probably just a protocol problem. Reported to work on zebrafish.

'''XAO:'''

References: Dev Bio 171:531- [http://www.xenbase.org/literature/article.do?method=display&articleId=19219]