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96768
TREM1 (D7D6I) Rabbit mAb (Flow Specific)
Primary Antibodies
Monoclonal Antibody

TREM1 (D7D6I) Rabbit mAb (Flow Specific) #96768

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Flow Cytometry Image 1 - TREM1 (D7D6I) Rabbit mAb (Flow Specific)

Flow cytometric analysis of human peripheral blood mononuclear cells, untreated (blue) or treated with LPS (300 ng/ml, 16 hr) #14011 (green), using TREM1 (D7D6I) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (dashed lines) co-stained with a CD14 antibody #36377. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody. Analysis was performed on CD14+ cells in the monocyte gate.

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Supporting Data

REACTIVITY H
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
Flow Cytometry 1:50

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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Flow Cytometry, Extracellular Epitope Protocol for Rabbit Antibodies

A. Solutions and Reagents

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

  1. 1X Phosphate Buffered Saline (PBS): To prepare 1 L 1X PBS: add 100 ml 10X PBS (#12528) to 900 ml water, mix.
  2. 4% Formaldehyde, Methanol-free (#47746), Optional
  3. Antibody Dilution Buffer: Purchase ready-to-use Flow Cytometry Antibody Dilution Buffer (#13616), or prepare a 0.5% BSA PBS buffer by dissolving 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.
  4. Recommended Anti-Rabbit secondary antibodies:

NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry.

NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.

B. Fixation

NOTE: If live cell staining is desired, proceed to Section C.

  1. Pellet cells by centrifugation and remove supernatant.
  2. Resuspend cells in approximately 100 µl 4% formaldehyde per 1 million cells. Mix well to dissociate pellet and prevent cross-linking of individual cells.
  3. Fix for 15 min at room temperature (20-25°C).
  4. Wash by centrifugation with excess 1X PBS. Discard supernatant in appropriate waste container. Resuspend cells in 1X PBS.

C. Immunostaining

NOTE: Count cells using a hemocytometer or alternative method.

  1. Aliquot desired number of cells into tubes or wells. (Generally, 5x105 to 1x106 cells per assay.)
  2. Pellet cells by centrifugation and remove supernatant.
  3. Resuspend cells in 100 µl of diluted primary antibody, prepared in Antibody Dilution Buffer at the recommended dilution.
  4. Incubate for 30-60 minutes at room temperature (20-25°C).
  5. Wash by centrifugation in Antibody Dilution Buffer. Discard supernatant. Repeat.
  6. Resuspend cells in 100 µl of diluted fluorochrome-conjugated secondary antibody, prepared in Antibody Dilution Buffer at the recommended dilution.
  7. Incubate for 30 minutes at room temperature.
  8. Wash by centrifugation in incubation buffer. Discard supernatant. Repeat.
  9. Resuspend cells in 200-500 µl of 1X PBS and analyze on flow cytometer.

posted January 2009

revised June 2020

Protocol Id: 133

Specificity / Sensitivity

TREM1 (D7D6I) Rabbit mAb recognizes endogenous levels of total TREM1 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the extracellular domain of human TREM1 protein.

Background

The triggering receptor expressed on myeloid cells 1 (TREM1) protein is an innate immune receptor that is primarily expressed on the cell surface of myeloid cells (1). TREM1 is a single-pass type I membrane glycoprotein that consists of an extracellular immunoglobulin-like domain, a transmembrane domain, and a cytoplasmic tail. TREM1, like its related protein TREM2, interacts with the tyrosine kinase-binding protein DAP12 to form a receptor-signaling complex (2). By accepting a diverse array of ligands, TREM1-expressing macrophages and neutrophils modulate inflammation through cytokine, chemokine, and receptor upregulation (2,3).

  1. Colonna, M. (2003) Nat Rev Immunol 3, 445-53.
  2. Thankam, F.G. et al. (2016) Expert Rev Clin Immunol 12, 1239-1249.
  3. Yuan, Z. et al. (2014) J Biol Chem 289, 15118-29.

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.