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10284
CD38 (HIT2) Mouse mAb (PerCP-Cy5.5® Conjugate)
Antibody Conjugates

CD38 (HIT2) Mouse mAb (PerCP-Cy5.5® Conjugate) #10284

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Flow cytometric analysis of live human peripheral blood mononuclear cells using CD38 (HIT2) Mouse mAb (PerCP-Cy5.5® Conjugate) (solid line) compared to concentration-matched Mouse (MOPC-21) mAb IgG1 Isotype Control (PerCP-Cy5.5® Conjugate) #24589 (dashed line).

To Purchase # 10284S
Product # Size Price
10284S
500 µl  (100 tests) $ 349

Supporting Data

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

Product Description

This Cell Signaling Technology antibody is conjugated to PerCP-Cy5.5® 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, pH 7.2. 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, Live Cell Protocol for Directly Conjugated 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. 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.

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/flowdyes for a listing of cellular dyes validated for use in flow cytometry.

B. Immunostaining

NOTE: Count cells using a hemocytometer or alternative method.

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

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.

  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 a recommended dilution or as determined via titration.
  4. Incubate for 30 min to 1 hr on ice. Protect from light.
  5. Wash by centrifugation in Antibody Dilution Buffer. Discard supernatant. Repeat.
  6. Resuspend cells in 200-500 µl of Antibody Dilution Buffer and analyze on flow cytometer.

posted June 2017

revised August 2019

Protocol Id: 1504

Specificity / Sensitivity

CD38 (HIT2) Mouse mAb (PerCP-Cy5.5® Conjugate) recognizes endogenous levels of total CD38 protein. This antibody detects epitopes 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

Cyclic ADP-ribose hydrolase 1 (CD38) is a transmembrane protein involved in several important biological processes, including immune response, insulin secretion, and social behavior. Originally described as a glycosylated immune cell surface marker, additional research determined that CD38 is a multifunctional enzyme that catalyzes the synthesis and hydrolysis of cyclic ADP ribose (cADPR) from NAD (1,2). Under acidic conditions, CD38 also catalyzes the synthesis of nicotinic acid adenine dinucleotide phosphate (NAADP) from NADP+. Both cADPR and NAADP act as calcium ion mobilizing messengers that target different intracellular Ca2+ stores (3-6). Since CD38 is the primary mammalian NAD+ glycohydrolase responsible for NAD+ metabolism, CD38 may be a valuable therapeutic target for treatment of metabolic diseases regulated by NAD+-dependent pathways (7,8). CD38 has also been considered a possible therapeutic target for antibody-mediated therapy for myeloma and chronic lymphocytic leukemia (9-11).

  1. Malavasi, F. et al. (2008) Physiol Rev 88, 841-86.
  2. Jin, D. et al. (2007) Nature 446, 41-5.
  3. Lee, H.C. et al. (1999) Mol Cell Biochem 193, 89-98.
  4. Calcraft, P.J. et al. (2009) Nature 459, 596-600.
  5. Ogunbayo, O.A. et al. (2011) J Biol Chem 286, 9136-40.
  6. Lee, H.C. (2012) J Biol Chem 287, 31633-40.
  7. Cantó, C. et al. (2012) Cell Metab 15, 838-47.
  8. Escande, C. et al. (2013) Diabetes 62, 1084-93.
  9. Malavasi, F. et al. (2011) Blood 118, 3470-8.
  10. Deaglio, S. et al. (2010) Semin Cancer Biol 20, 416-23.
  11. Chillemi, A. et al. (2013) Mol Med 19, 99-108.

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.