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90840
TMEM119 (E3E1O) Rabbit mAb
Primary Antibodies
Monoclonal Antibody

TMEM119 (E3E1O) Rabbit mAb #90840

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  1. IF

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).

To Purchase # 90840S
製品番号 サイズ 価格 在庫
90840S
100 µl

Supporting Data

REACTIVITY M
SENSITIVITY Endogenous
MW (kDa)
Source/Isotype Rabbit IgG

Application Key:

  • W-Western
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • IF-Immunofluorescence
  • F-Flow Cytometry
  • E-P-ELISA-Peptide

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • All-All Species Expected

Product Usage Information

Application Dilution
Immunofluorescence (Frozen) 1:200 - 1:400

Storage

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.

Protocol

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Immunofluorescence (Frozen)

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 serum / 0.3% Triton™ X-100): To prepare 10 ml, add 0.5 ml normal serum from the same species as the secondary antibody (e.g., Normal Goat Serum (#5425) to 9.5 ml 1X PBS) and mix well. 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. 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.

  6. Prolong® Gold AntiFade Reagent (#9071), Prolong® Gold AntiFade Reagent with DAPI (#8961).

B. Specimen Preparation - Frozen/Cryostat Sections (IF-F)

  1. For fixed frozen tissue proceed with Immunostaining (Section C).
  2. For fresh, unfixed frozen tissue, please fix immediately, as follows:
    1. Cover sections with 4% formaldehyde dilute in 1X PBS.

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

    2. Allow sections to fix for 15 minutes at room temperature.
    3. Aspirate liquid, rinse three times in 1X PBS for 5 minutes 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. Incubate specimen in fluorochrome-conjugated secondary antibody diluted in Antibody Dilution Buffer for 1–2 hr at room temperature in the dark.
  7. Rinse three times in 1X PBS for 5 min each.
  8. Coverslip slides with Prolong® Gold Antifade Reagent (#9071) or Prolong® Gold Antifade Reagent with DAPI (#8961).
  9. 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 July 2016

Protocol Id: 151

Specificity / Sensitivity

TMEM119 (E3E1O) Rabbit mAb recognizes endogenous levels of total TMEM119 protein.

Species Reactivity:

Mouse

Species predicted to react based on 100% sequence homology:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human TMEM119 protein.

Background

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).

  1. Bennett, M.L. et al. (2016) Proc Natl Acad Sci U S A 113, E1738-46.
  2. Satoh, J. et al. (2016) Neuropathology 36, 39-49.
  3. Deczkowska, A. et al. (2018) Cell 173, 1073-81.
  4. Keren-Shaul, H. et al. (2017) Cell 169, 1276-1290.e17.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
ProLong is a registered trademark of Life Technologies Corporation.

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