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96768
TREM1 (D7D6I) Rabbit mAb (Flow Specific)

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

APPLICATIONS

REACTIVITY SENSITIVITY MW (kDa) Isotype
H Endogenous Rabbit IgG
Flow Cytometry

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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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. 20X Phosphate Buffered Saline (PBS): (#9808) To prepare 1 L 1X PBS: add 50 ml 20X PBS to 950 ml dH2O, mix.
  2. Formaldehyde (methanol free).
  3. Incubation Buffer: Dissolve 0.5 g bovine serum albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.
  4. Recommended Anti-Rabbit secondary antibodies:

B. Fixation

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

  1. Collect cells by centrifugation and aspirate supernatant.
  2. Resuspend cells briefly in 0.5-1 ml PBS. Add formaldehyde to obtain a final concentration of 4% formaldehyde.
  3. Fix for 15 minutes at room temperature.
  4. Wash by centrifugation with excess 1X PBS. Discard supernatant in appropriate waste container. Resuspend cells in 1X PBS.
  5. Proceed with staining or store cells at +4°C in PBS with 0.1% sodium azide.

C. Immunostaining

NOTE: Count cells using a hemocytometer or alternative method.

  1. Aliquot desired number of cells into tubes or wells.
  2. If necessary, centrifuge to remove excess PBS.
  3. Resuspend cells in 100 µl of diluted primary antibody (prepared in incubation buffer at the recommended dilution).
  4. Incubate for 30-60 minutes at room temperature.
  5. Wash by centrifugation in incubation buffer. Discard supernatant. Repeat.
  6. Resuspend cells in fluorochrome-conjugated secondary antibody 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 1X PBS and analyze on flow cytometer; alternatively, for DNA staining, proceed to Section D.

D. Optional DNA Dye

  1. Resuspend cells in 0.5 ml of DNA dye (e.g. Propidium Iodide (PI)/RNase Staining Solution #4087).
  2. Incubate for at least 5 min at room temperature.
  3. Analyze cells in DNA staining solution on flow cytometer.

posted January 2009

revised June 2017

Protocol Id: 133

Application Dilutions
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.

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

Species Reactivity:

Human

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

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.
Entrez-Gene Id
54210
Swiss-Prot Acc.
Q9NP99
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.

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Product # Size Price
96768S
100 µl (10 western blots) $ 255.0