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41134
TMEM119 (E3E4T) Mouse mAb
Primary Antibodies
Monoclonal Antibody

TMEM119 (E3E4T) Mouse mAb #41134

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  1. IHC
Immunohistochemistry Image 1 - TMEM119 (E3E4T) Mouse mAb

Immunohistochemical analysis of paraffin-embedded normal human brain using TMEM119 (E3E4T) Mouse mAb.

Immunohistochemistry Image 2 - TMEM119 (E3E4T) Mouse mAb

Immunohistochemical analysis of paraffin-embedded human cerebellum (left) or colon carcinoma (right) using TMEM119 (E3E4T) Mouse mAb (top) or TMEM119 Rabbit mAb (bottom). These two antibodies detect independent, unique epitopes on human TMEM119. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.

Immunohistochemistry Image 3 - TMEM119 (E3E4T) Mouse mAb

Immunohistochemical analysis of paraffin-embedded normal human lung using TMEM119 (E3E4T) Mouse mAb.

Immunohistochemistry Image 4 - TMEM119 (E3E4T) Mouse mAb

Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using TMEM119 (E3E4T) Mouse mAb (left) compared to concentration-matched Mouse (E5Y6Q) mAb IgG2a Isotype Control #61656 (right).

Immunohistochemistry Image 5 - TMEM119 (E3E4T) Mouse mAb

Immunohistochemical analysis of paraffin-embedded human lymphoma using TMEM119 (E3E4T) Mouse mAb.

Immunohistochemistry Image 6 - TMEM119 (E3E4T) Mouse mAb

Immunohistochemical analysis of paraffin-embedded 293T cell pellet, untransfected (left) or TMEM119-transfected (right), using TMEM119 (E3E4T) Mouse mAb.

Immunohistochemistry Image 7 - TMEM119 (E3E4T) Mouse mAb

Immunohistochemical analysis of paraffin-embedded normal human testis using TMEM119 (E3E4T) Mouse mAb.

To Purchase # 41134S
製品番号 サイズ 価格 在庫
41134S
100 µl

Supporting Data

REACTIVITY H
SENSITIVITY Endogenous
MW (kDa)
Source/Isotype Mouse IgG2a

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
Immunohistochemistry (Paraffin) 1:200 - 1:800

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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Immunohistochemistry (Paraffin)

A. Solutions and Reagents

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

  1. Xylene.
  2. Ethanol, anhydrous denatured, histological grade (100% and 95%).
  3. Deionized water (dH2O).
  4. Hematoxylin (optional).
  5. Wash Buffer:
    1. 1X Tris Buffered Saline with Tween® 20 (TBST): To prepare 1L 1X TBST add 100 ml 10X Tris Buffered Saline with Tween® 20 (#9997) to 900 ml dH20, mix.
  6. SignalStain® Antibody Diluent (#8112).
  7. 1X Citrate Unmasking Solution: To prepare 250 mL of 1X citrate unmasking solution, dilute 25 ml of SignalStain® Citrate Unmasking Solution (10X) (#14746) with 225 mL of dH2O.
  8. 3% Hydrogen Peroxide: To prepare 100 ml, add 10 ml 30% H2O2 to 90 ml dH2O.
  9. Blocking Solution: TBST/5% Normal Goat Serum or 1X Animal-Free Blocking Solution.
    1. TBST/5% Normal Goat Serum: to 5 ml 1X TBST, add 250 µl Normal Goat Serum (#5425).
    2. 1X Animal-Free Blocking Solution: to 4 mL of dH2O add 1 ml of Animal-Free Blocking Solution (5X) (#15019).
  10. Detection System: SignalStain® Boost IHC Detection Reagents (HRP, Mouse #8125).
  11. Substrate: SignalStain® DAB Substrate Kit (#8059).
  12. Hematoxylin: Hematoxylin (#14166).
  13. Mounting Medium: SignalStain® Mounting Medium (#14177).

B. Deparaffinization/Rehydration

NOTE: Do not allow slides to dry at any time during this procedure.

  1. Deparaffinize/hydrate sections:
    1. Incubate sections in three washes of xylene for 5 min each.
    2. Incubate sections in two washes of 100% ethanol for 10 min each.
    3. Incubate sections in two washes of 95% ethanol for 10 min each.
  2. Wash sections two times in dH2O for 5 min each.

C. Antigen Unmasking

For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.

D. Staining

  1. Wash sections in dH2O three times for 5 min each.
  2. Incubate sections in 3% hydrogen peroxide for 10 min.
  3. Wash sections in dH2O two times for 5 min each.
  4. Wash sections in wash buffer for 5 min.
  5. Block each section with 100–400 µl of preferred blocking solution for 1 hr at room temperature.
  6. Remove blocking solution and add 100–400 µl primary antibody diluted in SignalStain® Antibody Diluent (#8112) to each section. Incubate overnight at 4°C.
  7. Equilibrate SignalStain® Boost Detection Reagent (HRP, Mouse #8125) to room temperature.
  8. Remove antibody solution and wash sections with wash buffer three times for 5 min each.
  9. Cover section with 1–3 drops SignalStain® Boost Detection Reagent (HRP, Mouse #8125) as needed. Incubate in a humidified chamber for 30 min at room temperature.
  10. Wash sections three times with wash buffer for 5 min each.
  11. Add 1 drop (30 µl) SignalStain® DAB Chromogen Concentrate to 1 ml SignalStain® DAB Diluent and mix well before use.
  12. Apply 100–400 µl SignalStain® DAB to each section and monitor closely. 1–10 min generally provides an acceptable staining intensity.
  13. Immerse slides in dH2O.
  14. If desired, counterstain sections with hematoxylin (#14166).
  15. Wash sections in dH2O two times for 5 min each.
  16. Dehydrate sections:
    1. Incubate sections in 95% ethanol two times for 10 sec each.
    2. Repeat in 100% ethanol, incubating sections two times for 10 sec each.
    3. Repeat in xylene, incubating sections two times for 10 sec each.
  17. Mount sections with coverslips and mounting medium (#14177).

DETECTION REAGENT/SUBSTRATE COMPATIBILITY
RECOMMENDED
DETECTION REAGENTS
SignalStain® Boost IHC Detection Reagent (HRP, Mouse) #8125 SignalStain® Boost IHC Detection Reagent (AP, Mouse) #31926
COMPATIBLE
CHROMOGEN
SignalStain® DAB Substrate Kit #8059 SignalStain® Vibrant Red Alkaline Phosphatase Substrate Kit #76713
SignalStain® Vivid Purple Peroxidase Substrate Kit #96632  

NOTE: Use of detection reagents other than those specified in this protocol may require further optimization of the primary antibody to account for the different sensitivities of the detection reagents.


posted February 2010

revised June 2020

Protocol Id: 280

Specificity / Sensitivity

TMEM119 (E3E4T) Mouse mAb recognizes endogenous levels of total TMEM119 protein.

Species Reactivity:

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.
SignalStain is a trademark of Cell Signaling Technology, Inc.
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