3G8- pronephros and otic vesicle and 4A6 – pronephros: Difference between pages

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'''Antigen:''' dissected pro nephroi, specific target unknown
'''Antigen:''' dissected pronephroi, target unknown
'''Xenbase Gene Page:'''
'''Xenbase Gene Page:'''
'''Epitope:'''
'''Epitope:''' mouse monoclonal IgG
'''Host Species/Isotype:''' mouse monoclonal IgG
'''Host Species/Isotype:'''
'''Antigen Species:'''
'''Antigen Species:''' IF, IP, WB
'''Applications:''' IF
'''Applications:''' 4A6
'''Antibody Name/Catalog Number:''' 3G8
'''Antibody Name/Catalog Number:''' 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.
'''Supplier:''' not sentitive to bleaching, does not function post in situ
'''Comments:''' IP unknown, does not work on Westerns
'''Comments:''' 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]
'''Contributor:'''
'''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.
'''Stains:''' proximal pronephric nephron, nephrostomes and otic vesicle,  
'''Additional Information:'''
'''Developmental Window:''' First detects target at stage 39, continues until latest stages tested.
''Stains:''' distal pronephric nephron and pronephric / Wolffian duct. The duct staining is mosaic and only some cells are recognized. Sains the entire cell surface evenly with no basolateral/apical bias.
'''Fixatives:''' MEMFA
''Available as:''' cell supernatant, 3 x
'''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.
'''Other species:''' Xenopus only
'''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
 
'''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:'''
'''Reference(s):''' Dev Bio 171:531- [http://www.xenbase.org/literature/article.do?method=display&articleId=19219]
applications abbreviations:
applications abbreviations:

Revision as of 13:04, 8 March 2011

Antigen: dissected pronephroi, target unknown

Xenbase Gene Page:

Epitope: mouse monoclonal IgG

Host Species/Isotype:

Antigen Species: IF, IP, WB

Applications: 4A6

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

Supplier: not sentitive to bleaching, does not function post in situ

Comments: Generated and characterized by Elizabeth Jones [1]. References: Dev Bio 171:531- [2]

Contributor:


Additional Information: Developmental Window: First detects target at stage 39, continues until latest stages tested. Stains:' distal pronephric nephron and pronephric / Wolffian duct. The duct staining is mosaic and only some cells are recognized. Sains the entire cell surface evenly with no basolateral/apical bias. Fixatives: MEMFA Available as:' cell supernatant, 3 x 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. Other species: Xenopus only

Reference(s): Dev Bio 171:531- [3]


applications abbreviations:

WB - Western blotting,

IP - immunoprecipitation,

IF - immunofluorescence,

IHC - immunohistochemistry,

EMSA - electromobility shift assay,

ChIP - chromatin immunoprecipitation,

FCM - flow cytometry

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