4A6; pronephric duct, pronephros

Antibody 4A6; pronephric duct

Generated by Liz Jones, University of Warwick, Coventry. Pictures by Peter Vize and Inga Dunkerson. Stains the pronephric duct from stage 38/39 onwards, and the nephrostomes. A full description of the generation and specificity of this antibody can be found in Vize, Jones and Pfister (1995) Developmental Biology 171; 531-540. To see Balinsky's figure of duct anatomy to compare with staining patterns click here.

A. The first stain is strongest in the most anterior region of the duct at stage 38. This is a little strange as the duct extends all the way to the cloaca and is quite functional by this stage. Single isolated stained cells are found at this stage amongst the unstained posterior portion of the duct.
B. and C. Closes up of similar stages.
D.

D. Close up of the posterior region of the duct adjacent to the cloaca, note the isolated 4A6 positive cells.
E. Right at the developing cloaca just three dispursed cells stain.
F.

F. Double stain of a similar stage showing the relationship of 4A6 duct staining (in red) to the pronephric tubules (3G8 stain) in blue.
G. Lateral close up view of stage 42, red stain.
H. A false color of this in case the red doesn't show up.
I.

I. Dorsal view of 4A6 stained duct.
J. The antigen is present on the surface of duct cells, you can see this under the stereoscope, it stains the entire surface evenly. Here is a wax cross section through a stained duct

Figures from the original 4A6 and 3G8 manuscript Developmental Biology 171; 531-540

Search PubMed for papers by Peter Vize or by Liz Jones
or for other papers using 4A6

Technical notes

Reference; Vize, Jones and Pfister (1995) Developmental Biology 171; 531-540
Antibody name; 4A6
Raised in; mouse, monoclonal
immunogen; dissected Xenopus pronephroi
epitope; unknown. 50 kD in size.
Subtype; IgG
fixatives; formalin, MEMFA, TCA are all OK. Others not tested.
immunoprecipitation/Western blotting?; i.p unknown, works on Westerns (50 kD band).
Available as; culture supernatent, 10 ml 2x stock. Shipped on dry ice so please supply account number for shipping costs.
Available from; Liz Jones ea@dna.bio.warwick.ac.uk.
Stock concentration; 2x
Optimized conditions; Works well with BCIP or red substrate. Development time typically about 30 minutes at room temp. Only note of caution, in some samples stain extends into pronephric tubules (for example), but it always looks fuzzy when this happens. This may be caused by disruption of cells during rehydration releasing the epitope from duct cells.
Species specificity; does not work on axolotl or zebrafish (I.Drummond, pers.comm.). Not tested further.

all of these images are 100% digital, and were captured directly from the microscope by a NeXTDimension computer system

peter@pvize.zo.utexas.edu

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