WB analysis of Tetrahymena themophila or Tetrahymena themophila expressing GFP-MacNup98A cell lysate using GTX00693 NUP98 antibody [13C2], GTX00695 NUP98 antibody [21A10], or GTX00697 NUP98 antibody [2H10].
Diamonds and asterisks represent uncharacterized proteins.

WB analysis of HeLa whole cell lysate using GTX00693 NUP98 antibody [13C2], GTX00695 NUP98 antibody [21A10], or GTX00697 NUP98 antibody [2H10]. Signals were detected using the same staining conditions, except that the exposure time of the middle lane (labeled 21A10 long exposure) was about 10 times longer than the exposure times of the other lanes.
Dilution : 0.4 μg/ml

WB analysis of S. pombe cell extracts using GTX00693 NUP98 antibody [13C2], GTX00695 NUP98 antibody [21A10], or GTX00697 NUP98 antibody [2H10]. Left and right lanes represent specimens from a wild type strain and an S. pombe strain in which Nup98 was chromosomally replaced with Nup98-GFP, respectively.
Dilution :
13C2 or 21A10 : 1:10
2H10 : 1 μg/ml

WB analysis of S. cereviciae cell extracts using GTX00693 NUP98 antibody [13C2], GTX00695 NUP98 antibody [21A10], or GTX00697 NUP98 antibody [2H10]. Arrows represent the positions of the indicated nucleoporins. Asterisks represent uncharacterized proteins.
Dilution :
13C2 or 21A10 : 1:10
2H10 : 20 μg/ml

ICC/IF analysis of Tetrahymena themophila cells using GTX00693 NUP98 antibody [13C2], GTX00695 NUP98 antibody [21A10], or GTX00697 NUP98 antibody [2H10]. The open arrow indicates the micronucleus. Insets are magnified images showing the position of the micronucleus. Both clone 13C2 and 21A10 stained the macronuclear periphery of T. thermophila. 13C2 mAb was highly specific to the macronucleus. In contrast, in addition to clear macronuclear staining, clone 21A10 also stained the micronuclear periphery. This indicates that 21A10 recognizes Nups localizing to the micronucleus such as Nup308 in addition to MacNup98A. Neither could 2H10 nor 414 could stain nuclear periphery of Tetrahymena.
Green : Primary antibody
Violet : DAPI
Dilution : 0.5 μg/ml
Fixation : Cold Methanol (-30 degree C) for 30 min

ICC/IF analysis of S. pombe cells using GTX00693 NUP98 antibody [13C2], GTX00695 NUP98 antibody [21A10], or GTX00697 NUP98 antibody [2H10].
Green : Primary antibody
Violet : DAPI
Fixation : 4% PFA for 10 min, treated with 0.6 mg/ml Zymolyase 100T at 3 degree C for 70 min
Permeabilization : 1% Triton X-100 for 1 min

ICC/IF analysis of HeLa cells using GTX00693 NUP98 antibody [13C2], GTX00695 NUP98 antibody [21A10], or GTX00697 NUP98 antibody [2H10]. The signal at the nuclear periphery with 21A10 mAb was much higher and the background lower than that of 2H10 and 13C2 antibodies.
Green : Primary antibody
Violet : DAPI
Dilution : 0.5 μg/ml
Fixation : Cold Methanol (-30 degree C) for 30 min

Summary of the suitability of GTX00693 NUP98 antibody [13C2], GTX00695 NUP98 antibody [21A10], or GTX00697 NUP98 antibody [2H10] for immunological applications.

ICC/IF analysis of S. cereviciae cells using GTX00693 NUP98 antibody [13C2], GTX00695 NUP98 antibody [21A10], or GTX00697 NUP98 antibody [2H10].
Green : Primary antibody
Violet : DAPI
Dilution :
13C2 or 21A10 : 1:10
2H10 : 10 μg/ml