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REACTIVITY SENSITIVITY MW (kDa) Isotype
H M R Mk Endogenous 98 Rabbit IgG
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Confocal immunofluorescent analysis of HeLa cells using NUP98 (C39A3) Rabbit mAb (Alex Fluor® 488 Conjugate) (green). Actin filaments were labeled with DyLight™ 554 Phalloidin (red).

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Immunofluorescence (Immunocytochemistry)

A. Solutions and Reagents

NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.

  1. 20X Phosphate Buffered Saline (PBS): (9808) To prepare 1L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix. Adjust pH to 8.0.
  2. Formaldehyde: 16%, methanol free, Polysciences, Inc. (cat# 18814), use fresh and store opened vials at 4°C in dark, dilute in 1X PBS for use.
  3. Blocking Buffer (1X PBS / 5% normal goat serum (#5425) / 0.3% Triton™ X-100): To prepare 10 ml: add 0.5 ml normal goat serum and 0.5 ml 20X PBS to 9.0 ml dH2O, mix. While stirring, add 30 µl Triton™ X-100.
  4. Antibody Dilution Buffer: (1X PBS / 1% BSA / 0.3% Triton™ X-100): To prepare 10 ml, add 30 µl Triton™ X-100 to 10 ml 1X PBS. Mix well then add 0.1 g BSA (#9998), mix.
  5. Prolong® Gold AntiFade Reagent (#9071), Prolong® Gold AntiFade Reagent with DAPI (#8961).

B. Specimen Preparation - Cultured Cell Lines (IF-IC)

NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.

  1. Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.

    NOTE: Formaldehyde is toxic, use only in a fume hood.

  2. Allow cells to fix for 15 min at room temperature.
  3. Aspirate fixative, rinse three times in 1X PBS for 5 min each.
  4. Proceed with Immunostaining (Section C).

C. Immunostaining

NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.

  1. Block specimen in Blocking Buffer for 60 min.
  2. While blocking, prepare primary antibody by diluting as indicated on datasheet in Antibody Dilution Buffer.
  3. Aspirate blocking solution, apply diluted primary antibody.
  4. Incubate overnight at 4°C.
  5. Rinse three times in 1X PBS for 5 min each.
  6. Coverslip slides with Prolong® Gold Antifade Reagent (#9071) or Prolong® Gold Antifade Reagent with DAPI (#8961).
  7. For best results, allow mountant to cure overnight at room temperature. For long-term storage, store slides flat at 4°C protected from light.

posted November 2006

revised November 2013

protocol id: 182

Product Usage Information

Application Dilutions
Immunofluorescence 1:50

Storage: Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

Specificity / Sensitivity

NUP98 (C39A3) Rabbit mAb (Alexa Fluor® 488 Conjugate) detects endogenous levels of total NUP98 protein.


Species Reactivity: Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro671 of human NUP98.

Product Description

This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct immunofluorescence analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated NUP98 (C39A3) Rabbit mAb # 2598.


Nucleoporin 98 kDa (NUP98) is a component of the nuclear pore complex. It is expressed as three different precursors that undergo auto-cleavage to generate a common amino-terminal 98 kDa peptide (NUP98) and carboxy-terminal 6, 96 (NUP96) and 88 (p88) kDa peptides (1,2). NUP98 contains FG and GLFG repeat domains at its amino terminus and a RNA-binding domain in its carboxy terminus (3). The NUP98 gene is localized on chromosome 11p15.5, a region frequently rearranged in leukemias. To date, 15 fusion partners have been identified for NUP98 (4,5).


1.  Fontoura, B.M. et al. (1999) J Cell Biol 144, 1097-112.

2.  Rosenblum, J.S. and Blobel, G. (1999) Proc Natl Acad Sci U S A 96, 11370-5.

3.  Griffis, E.R. et al. (2004) Mol Biol Cell 15, 1991-2002.

4.  Slape, C. and Aplan, P.D. (2004) Leuk Lymphoma 45, 1341-50.

5.  Nakamura, T. (2005) Int J Hematol 82, 21-7.


Entrez-Gene Id 4928
Swiss-Prot Acc. P52948


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor® is a registered trademark of Life Technologies Corporation.
The Alexa Fluor® dye antibody conjugates in this product are sold under license from Life Technologies Corporation for research use only, except for use in combination with DNA microarrays. The Alexa Fluor® dyes (except for Alexa Fluor® 430 dye) are covered by pending and issued patents. Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
U.S. Patent No. 5,675,063.