Our anti-human (KELE) NIS antibody is a polyclonal affinity-purified antibody that detects both the native and denatured forms of the human sodium iodide symporter (hNIS). This antibody recognizes an intracellular C-terminal epitope. It does not detect mouse or rat NIS.

Immunohistochemistry staining for human NIS expression

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Recommended dilution: 1:5000

Thyroid tissue sections were deparaffinized. Slides were subjected to antigen retrieval using 10% citrate buffer. Slides were incubated with 1 ug/ml anti-human NIS antibody for 1h. All slides were counterstained with hematoxylin. A. normal thyroid. C. Graves’ tissue. F. papillary carcinoma. K. follicular carcinoma.

(performed by Dr. Nancy Carrasco’s laboratory in Dohan et al. 2001)

Immunoblotting for human NIS protein

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Recommended dilution: 1:1000 to 1:5000

Stimulation of NIS protein expression and plasma membrane targeting by all-trans-retinoic acid (tRa) and hydrocortisone was determined by Dohan et al. 2006.

MCF-7 human breast cancer cells were treated for 48h with trans-retinoic acid (tRa) or hydrocortisone (H). Cells were lyzed and subjected to immunoblot analysis with 4nM anti-hNIS antibody. A 100 kDa band of mature NIS and 60 kDa band of partially glycoslyated polypeptide were detected.

For western blotting of NIS proteins, it is recommended that samples be heated at 37°C for 30 minutes prior to loading for SDS-PAGE (do not boil).

Immunofluorescence

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Recommended dilution: 1:500

NIS targeting to plasma membrane was assessed by immunoblotting with anti-hNIS antibody. Cells were permeabilized with 0.2% BSA, 0.1% triton X-100 in PBS for 10min and then quenched with 50 mM NH4Cl in PBS for 10 min. Cells were incubated with 4nM anti-hNIS antibody for 1h at room temperature. Cells were washed and incubated with anti-rabbit FITC conjugated secondary antibody. Nontreated MCF-7 (G); intracellular and faint plasma membrane-localized NIS expression in tRa-treated MCF-7 (H); clear plasma membrane localization of NIS in tRaH-treated MCF-7 (I).

Note: This antibody recognizes the cytosolic C-terminus of hNIS. Therefore, samples must be permeabilized prior to incubation with anti-human NIS antibody for immunohistochemistry and immunofluorescence.

Product Citations

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Carrasco et al. Methods Enzymol. 1986. 125:453-467

Dohan et al. Mol  Endoclinol. 2006.  20:1121-1137

Dohan et al. J Clin Endocrinol Metab. 2001. 86:2697-270

Tazebay et al. Nat. Med. 2000. 6(8):871-8