CD38 Antibody, anti-human, REAfinity™

Name: CD38 Antibody, anti-human, REAfinity™
Availability: InStock

CD38 Antibody, anti-human, REAfinity™Extended ValidationRecombinant

Clone:

REA572

Type of antibody:

Primary antibodies, Recombinant antibodies

Isotype:

recombinant human IgG1

Applications:

FC, MICS, IF, IHC, MC

Alternative names:

ADPRC 1, ADPRC1, T10

Flow cytometry applications forCD38 Antibody, anti-human, REAfinity™

APC
Biotin
PE
PE-Vio 615
PE-Vio 770
Vio Bright FITC
Vio Bright R720
Vio Bright V423

Conjugate:

APC

Recommended dilution:

1:50

Remark:

Cells should be stained prior to fixation, if formaldehyde is used as a fixative.

Tested:

QC tested

Products used:

130-113-991, 130-113-429

Protocols:

Cell surface flow cytometry staining protocol (PBS/EDTA/BSA 1:50)

Description:

Human peripheral blood mononuclear cells (PBMCs) were stained with CD38 antibodies or with the corresponding REA Control (S) antibodies (left images) as well as with CD14 antibodies. Flow cytometry was performed using the MACSQuant^® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

MICS (MACSima Imaging Cyclic Staining) applications forCD38 Antibody, anti-human, REAfinity™

APC
PE

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

PFA

Tested:

during development

Species tested:

human

Products used:

130-113-991, 130-113-429

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Conjugate:

APC

Recommended dilution:

1:50

Fixation:

Acetone

Tested:

during development

Species tested:

human

Products used:

130-113-991, 130-113-429

Compatible applications:

IF, IHC

Protocols:

Protocol for MICS experiments

Mass cytometry applications forCD38 Antibody, anti-human, REAfinity™

pure

Conjugate:

pure

Products used:

130-122-307

Extended validation for CD38 Antibody, anti-human, REAfinity™

Specificity

Epitope competition

In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.

Other clones	Overlap in epitope recognition with REA572
HB7	++
HIT2	++
IB6	++
REA671	++
Cells were incubated with an excess of purified unconjugated CD38 (REA572) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Knockout validation

To ensure antibody specificity, the target gene is knocked out in a suitable cell line using the CRISPR/Cas9 system and the knockout is confirmed by sequencing of the target locus. The antibody is considered to bind specifically to the intended epitope if no antibody binding to the knockout cells can be detected. The antibody staining is controlled by fluorescence microscopy and/or flow cytometry.

WT

KO

Fluorescence microscopy image of CD38 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD38-PE (REA572, red) and counterstained with DRAQ5 (blue) as DNA stain.

Fluorescence microscopy image of CD38 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD38-PE (REA572, red) and counterstained with DRAQ5 (blue) as DNA stain.

Overlay histogram showing flow cytometric analysis of CD38 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD38-PE, clone (REA572). Flow cytometry was performed with the MACSQuant

^®

Analyzer. Cell debris, dead cells and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Overlay histogram showing flow cytometric analysis of CD38 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD38-PE, clone (REA572). Flow cytometry was performed with the MACSQuant

^®

Analyzer. Cell debris, dead cells and cell doublets were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Sensitivity

Performance comparison

Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.

Flow cytometric comparison of different clones for CD38. Human peripheral blood mononuclear cells (PBMCs) were stained with CD38antibodies and with a suitable counterstaining. As a control, CD38 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD38. Human peripheral blood mononuclear cells (PBMCs) were stained with CD38antibodies and with a suitable counterstaining. As a control, CD38 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD38. Human peripheral blood mononuclear cells (PBMCs) were stained with CD38antibodies and with a suitable counterstaining. As a control, CD38 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD38. Human peripheral blood mononuclear cells (PBMCs) were stained with CD38antibodies and with a suitable counterstaining. As a control, CD38 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD38. Human peripheral blood mononuclear cells (PBMCs) were stained with CD38antibodies and with a suitable counterstaining. As a control, CD38 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Flow cytometric comparison of different clones for CD38. Human peripheral blood mononuclear cells (PBMCs) were stained with CD38antibodies and with a suitable counterstaining. As a control, CD38 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant^® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Fixation data

To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.

No fixation

Post fixation

Comparison of staining pattern on non-fixed and fixed cells using CD38 (REA572). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Comparison of staining pattern on non-fixed and fixed cells using CD38 (REA572). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD38 Antibody, anti-human, REAfinity™

Overview

Clone REA572 recognizes the human CD38 antigen, a single-chain type II transmembrane glycoprotein with enzymatic activity. It is present on the majority of hematopoietic cells, prevalent during early differentiation and activation processes. Terminally differentiated B cells (plasma cells) express CD38 brightly. Furthermore, CD38 is constitutively expressed in several tissues, for example brain, muscle, and kidney. CD38, a disease marker for human leukemias and myelomas, plays a role in the pathogenesis and outcome of human immunodeficiency virus infection and chronic lymphocytic leukemia, and controls insulin release and also the development of diabetes. Furthermore it catalyzes the synthesis and hydrolysis of cyclic ADP-ribose (cADPR) from NAD

⁺

to ADP-ribose which is essential for the regulation of intracellular Ca

²⁺

.

Additional information: Clone REA572 displays negligible binding to Fc receptors.

Alternative names

ADPRC 1, ADPRC1, T10

Detailed product information

Technical specifications

Clone	REA572
Clonality	monoclonal
Isotype	recombinant human IgG1
Isotype control	REA Control Antibody (S), human IgG1
Host	human cell line
Type of antibody	Primary antibodies, Recombinant antibodies
Species	human
Antigen	CD38
Alternative names of antigen	ADPRC 1, ADPRC1, T10
Molecular mass of antigen [kDa]	34
Distribution of antigen	B cells, leukocytes, lymphocytes, monocytes, myeloid leukemia cells, NK cells, red blood cells, T cells, basophils, pancreatic carcinoma cells, plasma cells, thymocytes, bone marrow, brain, kidney, placenta, ovary, skeletal muscle
Entrez Gene ID	952
RRID	AB_2726165, AB_2733808, AB_2733809, AB_2726441, AB_2726164, AB_2733227, AB_2733228, AB_2751179, AB_2751112, AB_2801888, AB_2889570, AB_2889569, AB_2904978, AB_2904977, AB_2921873, AB_2921860, AB_2726442

Resources for CD38 Antibody, anti-human, REAfinity™

Certificates

Please follow this

link

to search for Certificates of Analysis (CoA) by lot number.

References for CD38 Antibody, anti-human, REAfinity™

Publications

Jackson, D. G. and Bell, J. I. (1990) Isolation of a cDNA encoding the human CD38 (T10) molecule, a cell surface glycoprotein with an unusual discontinuous pattern of expression during lymphocyte differentiation. J Immunol 144(7): 2811-2815
Sathish, V. et al. (2014) Inflammation, caveolae and CD38-mediated calcium regulation in human airway smooth muscle. Biochim. Biophys. Acta 1843(2): 346-351
Malavasi, F. et al. (2008) Evolution and function of the ADP ribosyl cyclase/CD38 gene family in physiology and pathology. Physiol. Rev. 88(3): 841-886

Applications

Extended validation