Lane 1 : Anti-NUP98 antibody [13C2 + 21A10] - BSA and Azide free (ab179911) at 0.4 µg/ml (13C2 clone)
Lane 2 : Anti-NUP98 antibody [13C2 + 21A10] - BSA and Azide free (ab179911) at 0.4 µg/ml (21A10 clone)

All lanes : HeLa cell extract

Secondary
All lanes : HRP-labeled anti-mouse IgG at 0.4 µg/ml

Developed using the ECL technique.

Predicted band size: 112 kDa

Immunofluorescence analysis of methanol-fixed HeLa cells, labeling NUP98 using two different clones of ab179911 at 0.5 µg/ml, followed by Alexa Fluor 488-conjugated anti-mouse lgG (green) at 4 µg/ml. DAPI was used to stain DNA (magenta). Upper and middle panels correspond to black-and-white images while the bottom panel represents merged colored images.

Immunofluorescence analysis of methanol-fixed Tetrahymena thermophila cells, labeling NUP98 using two different clones of ab179911 at 0.5 µg/ml, followed by Alexa Fluor 488-conjugated anti-mouse lgG (green) at 4 µg/ml. DAPI was used to stain DNA (magenta). Upper and middle panels correspond to black-and-white images while the bottom panel represents merged images. Dotted lines represent the outlines of cells. The open arrow indicates the micronucleus. Insets are magnified images showing the position of the micronucleus.

Lanes 1 & 4 : Anti-NUP98 antibody [13C2 + 21A10] - BSA and Azide free (ab179911) at 2 µg/ml (13C2 clone)
Lanes 2-3 : Anti-NUP98 antibody [13C2 + 21A10] - BSA and Azide free (ab179911) at 2 µg/ml (21A10 clone)

Lanes 1-3 : Tetrahymena thermophila cell extract
Lane 4 : Cell extract from Tetrahymena thermophila expressing endogenously NUP98 fused to a fluorescence protein

Developed using the ECL technique.

Predicted band size: 112 kDa
Observed band size: 98 kDa why is the actual band size different from the predicted?
Additional bands at: 125 kDa (possible tagged protein)

Open arrowheads, wild type NUP98. Solid arrow, NUP98 fused to a fluorescence protein. Diamonds and asterisks represent uncharacterized proteins. For lane 3 exposure time was about 10 times longer than for the other lanes.

Immunofluorescence analysis of formaldehyde-fixed, zymolyase-treated, S. pombe cells, labeling NUP98 using two different clones of ab179911 at 10 µg/ml, followed by Alexa Fluor 488-conjugated anti-mouse lgG (green). DAPI was used to stain DNA (magenta). Upper and middle panels correspond to black-and-white images while the bottom panel represents merged images. Dotted lines represent the outlines of cells.

Lanes 1-2 : Anti-NUP98 antibody [13C2 + 21A10] - BSA and Azide free (ab179911) at 2 µg/ml (13C2 clone)
Lanes 3-4 : Anti-NUP98 antibody [13C2 + 21A10] - BSA and Azide free (ab179911) at 2 µg/ml (21A10 clone)

Lanes 1 & 3 : Cell extract from S. pombe wild type
Lanes 2 & 4 : Cell extract from S. pombe expressing endogenously NUP98 fused to a fluorescence protein

Developed using the ECL technique.

Predicted band size: 112 kDa

Immunofluorescence analysis of formaldehyde-fixed, zymolyase-treated, S. cerevisiae cells, labeling NUP98 using two different clones of ab179911 at 10 µg/ml, followed by Alexa Fluor 488-conjugated anti-mouse lgG (green). DAPI was used to stain DNA (magenta). Upper and middle panels correspond to black-and-white images while the bottom panel represents merged images. Dotted lines represent the outlines of cells.

Lane 1 : Anti-NUP98 antibody [13C2 + 21A10] - BSA and Azide free (ab179911) at 2 µg/ml (13C2 clone)
Lane 2 : Anti-NUP98 antibody [13C2 + 21A10] - BSA and Azide free (ab179911) at 2 µg/ml (21A10 clone)

All lanes : S. cerevisiae cell extract

Developed using the ECL technique.

Predicted band size: 112 kDa

Asterisks represent uncharacterized proteins.

Summary of the suitability of ab179911 clones for immunological applications. IF: indirect immunofluorescence staining; WB: Western blotting analysis.