Confocal immunofluorescent analysis of wild-type mouse brain (left) and liver (right) using TMEM119 (E3E1O) Rabbit mAb (green). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Confocal immunofluorescent analysis of brain from the 5XFAD mouse model of Alzheimer’s disease. Sections were labeled with TMEM119 (E3E1O) Rabbit mAb (green) and GFAP (GA5) Mouse mAb #3670 (yellow). Plaques were then stained with β-Amyloid (D54D2) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #42284 (red) after blocking free secondary binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900. Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
|Immunofluorescence (Frozen)||1:200 - 1:400|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Recommended Fluorochrome-conjugated Anti-Rabbit secondary antibodies:
NOTE: When using any primary or fluorochrome-conjugated secondary antibody for the first time, titrate the antibody to determine which dilution allows for the strongest specific signal with the least background for your sample.
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 July 2016
Protocol Id: 151
TMEM119 (E3E1O) Rabbit mAb recognizes endogenous levels of total TMEM119 protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human TMEM119 protein.
Transmembrane protein 119 (TMEM119) is a cell-surface protein of unknown function, expressed exclusively by the microglia subset of myeloid and neural cells (1). Iba1+ microglia with both ramified and amoeboid morphologies express TMEM119, while Iba1+ macrophages are TMEM119 negative (2). TMEM119 and other homeostatic genes have been shown to be downregulated in disease-associated microglia (DAM). These DAM microglia are conserved in human and mouse and have been identified in Alzheimer’s disease (AD), amyotrophic lateral sclerosis (ALS), and 5XFAD and APP/PS1 mouse models (3,4). This protein’s specificity as a microglia marker has proven itself important for labeling microglia in healthy tissue as well as deciphering them from infiltrating macrophages and other cells types in neurodegenerative disease models (1,2).
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