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66423S 100 µl (50 tests) $299.00.0
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REACTIVITY SENSITIVITY MW (kDa) Isotype
H Endogenous Rabbit IgG
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Flow Cytometry

Flow cytometric analysis of human peripheral blood monocytes co-stained with anti-human CD14-eFluor® 450 antibody using Dectin-1 (E1X3Z) Rabbit mAb (PE Conjugate) (right) compared to a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (PE Conjugate) #5742 (left).

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Flow Cytometry, Methanol Permeabilization Protocol for Direct Conjugates

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. 16% Formaldehyde (methanol free).
  3. 100% methanol.
  4. Incubation Buffer: Dissolve 0.5 g Bovine Serum Albumin (BSA) (#9998) in 100 ml 1X PBS. Store at 4°C.

B. Fixation

NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to fixation.

  1. Collect cells by centrifugation and aspirate supernatant.
  2. Resuspend cells in 0.5-1 ml 1X PBS. Add formaldehyde to obtain a final concentration of 4%.
  3. Fix for 15 min at room temperature.
  4. Wash by centrifugation with excess 1X PBS. Discard supernatant in appropriate waste container. Resuspend cells in 0.5-1 ml 1X PBS.

C. Permeabilization

  1. Permeabilize cells by adding ice-cold 100% methanol slowly to pre-chilled cells, while gently vortexing, to a final concentration of 90% methanol.
  2. Incubate 30 min on ice.
  3. Proceed with immunostaining (Section D) or store cells at -20°C in 90% methanol.

D. Immunostaining

  1. Aliquot desired number of cells into tubes or wells.
  2. Wash cells by centrifugation in excess 1X PBS to remove methanol. Discard supernatant in appropriate waste container. Repeat if necessary.
  3. Resuspend cells in 100 µl of diluted antibody conjugate (prepared in incubation buffer at the recommended dilution).
  4. Incubate for 1 hr at room temperature. Protect from light.
  5. Wash by centrifugation in incubation buffer. Discard supernatant. Repeat.
  6. Resuspend cells in 1X PBS and analyze on flow cytometer; alternatively, for DNA staining, proceed to optional DNA stain (Section E).

E. 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 July 2009

revised June 2017

Protocol Id: 407

Product Usage Information

Application Dilutions
Flow Cytometry 1:50

Storage: Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.

Specificity / Sensitivity

Dectin-1 (E1X3Z) Rabbit mAb (PE Conjugate) recognizes endogenous levels of total Dectin-1 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 Dectin-1 protein.

Product Description

This Cell Signaling Technology antibody is conjugated to phycoerythrin (PE) and tested in-house for direct flow cytometry analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Dectin-1 (E1X3Z) Rabbit mAb #60128.


Dectin-1 is a C-type lectin receptor expressed by macrophages, monocytes, dendrtic cells, neutrophils, and a subset of γδ T cells (1,2). Dectin-1 is a glycoprotein with eight different isoforms, generated through alternative splicing (3-5). It plays an important role in anti-fungal immunity by acting as a pattern recognition receptor for β-glucans found on the cell wall of fungi and some bacteria (5,6). Dectin-1 is composed of a short amino-terminal cytoplasmic domain containing an ITAM-like motif, a transmembrane domain, and an extracellular carboxy-terminal C-type lectin domain (5). Dectin-1 recognizes β-glucans through its C-type lectin domain and transduces signals through its ITAM-like motif by recruiting and activating Syk (7,8). Dendritic cells activated through Dectin-1 promote differentiation of Th17 cells by producing IL-6 and IL-23 (9).


1.  Taylor, P.R. et al. (2002) J Immunol 169, 3876-82.

2.  Martin, B. et al. (2009) Immunity 31, 321-30.

3.  Ariizumi, K. et al. (2000) J Biol Chem 275, 20157-67.

4.  Kato, Y. et al. (2006) Biol Pharm Bull 29, 1580-6.

5.  Willment, J.A. et al. (2001) J Biol Chem 276, 43818-23.

6.  Brown, G.D. and Gordon, S. (2001) Nature 413, 36-7.

7.  Rogers, N.C. et al. (2005) Immunity 22, 507-17.

8.  Underhill, D.M. et al. (2005) Blood 106, 2543-50.

9.  LeibundGut-Landmann, S. et al. (2007) Nat Immunol 8, 630-8.


Entrez-Gene Id 64581
Swiss-Prot Acc. Q9BXN2


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.
eFluor is a registered trademark of eBioscience.

66423
Dectin-1 (E1X3Z) Rabbit mAb (PE Conjugate)