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90662S 100 µl (50 tests) $329.00.0
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REACTIVITY SENSITIVITY MW (kDa) Isotype
H Endogenous Rabbit IgG
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Flow Cytometry

Flow cytometric analysis of MUTZ-3 cells, undifferentiated (blue) or differentiated (green) into Langerhans-like cells by treatment with Human Granulocyte Macrophage Colony Stimulating Factor (hGM-CSF) #8922 (10ng/ml), Human Transforming Growth Factor β1 (hTGF-β1) #8915 (10ng/ml), and Human Tumor Necrosis Factor-α (hTNF-α) #8902 (2.5ng/ml) for 10 days, using Langerin (D9H7R) XP® Rabbit mAb (PE Conjugate).

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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

Langerin (D9H7R) XP® Rabbit mAb (PE Conjugate) recognizes endogenous levels of total Langerin protein.


Species Reactivity: Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val14 of human langerin 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 Langerin (D9H7R) XP® Rabbit mAb #13650.


Langerin (CD207) is a C-type lectin receptor whose expression is restricted mainly to dendritic cells in the skin including Langerhans cells in the epidermis and langerin+/CD103+ dermal dendritic cells (1-4). Langerin is found on the cell surface and within rod-shaped organelles called Birbeck granules, and its expression is required for the formation of Birbeck granules (1,5). Langerin recognizes carbohydrate motifs on the surface of pathogens, resulting in endocytosis of the pathogen into Birbeck granules, degradation of the pathogen, and antigen presentation to T cells (6-8).


1.  Valladeau, J. et al. (2000) Immunity 12, 71-81.

2.  Poulin, L.F. et al. (2007) J Exp Med 204, 3119-31.

3.  Ginhoux, F. et al. (2007) J Exp Med 204, 3133-46.

4.  Bursch, L.S. et al. (2007) J Exp Med 204, 3147-56.

5.  Kissenpfennig, A. et al. (2005) Mol Cell Biol 25, 88-99.

6.  Stambach, N.S. and Taylor, M.E. (2003) Glycobiology 13, 401-10.

7.  Idoyaga, J. et al. (2008) J Immunol 180, 3647-50.

8.  Igyártó, B.Z. et al. (2011) Immunity 35, 260-72.


Entrez-Gene Id 50489
Swiss-Prot Acc. Q9UJ71


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.

90662
Langerin (D9H7R) XP® Rabbit mAb (PE Conjugate)