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19805
CD27 (O323) Mouse mAb (APC-Cy7® Conjugate)
Antibody Conjugates

Monoclonal Antibody - CD27 (O323) Mouse mAb (APC-Cy7® Conjugate), UniProt ID P26842, Entrez ID 939 #19805

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Flow cytometric analysis of live human peripheral blood mononuclear cells using CD27 (O323) Mouse mAb (PE-Cy7® Conjugate) (solid line) compared to concentration-matched Mouse (MOPC-21) mAb IgG1 Isotype Control (PE-Cy7® Conjugate) #79339 (dashed line).

To Purchase # 19805S

Supporting Data

REACTIVITY H
SENSITIVITY Endogenous
MW (kDa)
Isotype Mouse IgG1, kappa

Product Description

This Cell Signaling Technology antibody is conjugated to PE-Cy7® and tested in-house for direct flow cytometric analysis in human cells.

Product Usage Information

Application Dilutions
Flow Cytometry 1:20

Storage:

Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH7.2. This product is stable for 6 months when stored at 4ºC. Do not aliquot the antibody. Protect from light. Do not freeze.

Protocol

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Flow Cytometry

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

B. Immunostaining

  1. Count cells using a hemacytometer or alternative method.
  2. Aliquot 0.5-1x106 cells into each assay tube (by volume).
  3. Add 2-3 ml Incubation Buffer to each tube and rinse by centrifugation. Repeat.
  4. Resuspend cells in 100 µl of diluted primary antibody (prepared in incubation buffer at the recommended dilution).
  5. Incubate for 1 hr at room temperature.
  6. Wash by centrifugation in 2-3 ml incubation buffer.
  7. Resuspend cells in the appropriate volume of PBS and analyze on flow cytometer.

posted June 2017

Protocol Id: 1504

Specificity / Sensitivity

CD27 (O323) Mouse mAb (PE-Cy7® Conjugate) recognizes endogenous levels of total CD27 protein. This antibody detects an epitope within the extracellular domain.

Species Reactivity:

Human

Source / Purification

This monoclonal antibody was purified from tissue culture supernatant via affinity chromatography. The purified antibody was conjugated under optimal conditions, with unreacted dye removed from the preparation.

Background

CD27 (TNFRSF7) is a transmemebrane protein of the TNF receptor superfamily (TNFRSF). It is mainly expressed on lymphoid cells (also on early hematopoietic precursor cells in mice) (1,2). CD27 is considered a phenotypic marker for memory B cells and is also used to identify B regulatory (Breg) cells (3,4). It is constitutively expressed on naïve CD4 and CD8 T cells and its expression is further upregulated upon T cell activation. CD27 is one of the two most important co-stimulatory receptors for T cell priming (the other one is CD28). While CD28 co-stimulatory signal mainly triggers cell proliferation, CD27 co-stimulatory signal primarily promotes cell survival and differentiation (5,6). Upon binding to its ligand CD70, CD27 activates the NF-κB and JNK signaling pathways through TNFR associated factors (TRAFs), the adaptor molecules that are associated with CD27 cytoplasmic tail domain. Upon activation CD27 is shed from cell surface and soluble CD27 is used as a marker of T cell activation (7,8).

  1. So, T. et al. (2006) Int J Hematol 83, 1-11.
  2. Waight, J.D. et al. (2017) Hum Antibodies 25, 87-109.
  3. Agematsu, K. (2000) Histol Histopathol 15, 573-6.
  4. Bouaziz, J.D. et al. (2012) Curr Mol Med 12, 519-27.
  5. Croft, M. Cytokine Growth Factor Rev 14, 265-73.
  6. Acuto, O. and Michel, F. (2003) Nat Rev Immunol 3, 939-51.
  7. Lens, S.M. et al. (1998) Semin Immunol 10, 491-9.
  8. van de Ven, K. and Borst, J. (2015) Immunotherapy 7, 655-67.

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.
Cy and CyDye are registered trademarks of GE Healthcare.