Confocal immunofluorescent analysis of rat testis (left, positive) or small intestine (right, negative) using NUT (C52B1) Rabbit mAb (Alexa Fluor® 555 Conjugate) (yellow). Actin filaments were labeled with DyLight™ 650 Phalloidin #12956 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 555 fluorescent dye and tested in-house for direct immunofluorescent analysis in rat tissue. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated NUT (C52B1) Rabbit mAb #3625.
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.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Cover sections with 4% formaldehyde dilute in 1X PBS.
NOTE: Formaldehyde is toxic, use only in fume hood.
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.
posted November 2006
revised November 2013
Protocol Id: 222
NUT (C52B1) Rabbit mAb (Alexa Fluor® 555 Conjugate) detects endogenous levels of total NUT protein. The antibody also detects endogenous levels of the BRD4-NUT fusion protein found in NUT midline carcinoma (NMC).
Monoclonal antibody is produced by immunizing animals with a recombinant protein corresponding to the human NUT protein.
Nuclear protein in testis (NUT) is normally confined to the germ cells of the testis and ovary (1,2). NUT midline carcinoma (NMC) is a recently recognized cancer that is defined by the presence of chromosomal rearrangements involving the NUT gene on chromosome 15q14 (3). In most cases the chromosomal translocation occurs between NUT and BRD4 on chromosome 19, resulting in the formation of a BRD4-NUT fusion protein. In the remaining tumors, variant NUT rearrangements are present involving BRD3, a very close homolog of BRD4. BRD4-NUT and BRD3-NUT encode fusion proteins that appear to contribute to carcinogenesis by blocking epithelial cell differentiation. NMCs, which are aggressive and highly lethal carcinomas, are morphologically indistinguishable from other poorly differentiated carcinomas. Given the limited expression of endogenous NUT protein, this antibody can be used to detect NUT fusion proteins in tissues by immunohistochemistry and immunofluorescence (2).
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