Clone:
REA671
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC, MC
Alternative names:
T10, ADPRC 1

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 clonesOverlap in epitope recognition with REA671
HB7++
HIT2++
IB6++
REA572++
Cells were incubated with an excess of purified unconjugated CD38 (REA671) 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
View details
Fluorescence microscopy image of CD38 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD38-PE (REA671, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Fluorescence microscopy image of CD38 knockout cells. Wild type (WT, left) and knockout cells (KO, right) were stained with CD38-PE (REA671, 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 (REA671, red) and counterstained with DRAQ5 (blue) as DNA stain.
View details
Overlay histogram showing flow cytometric analysis of CD38 knockout cells. Wild type (red) and knockout cells (blue) were stained with CD38-PE, clone (REA671). 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 (REA671). 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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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.
View details
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
View details
Comparison of staining pattern on non-fixed and fixed cells using CD38 (REA671). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD38 (REA671). 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 (REA671). 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 REA671 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 REA671 displays negligible binding to Fc receptors.

Alternative names

T10, ADPRC 1

Detailed product information

Technical specifications

CloneREA671
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD38
Alternative names of antigenT10, ADPRC 1
Molecular mass of antigen [kDa]34
Distribution of antigenB cells, leukocytes, lymphocytes, monocytes, myeloid 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 ID952
RRIDAB_2733811, AB_2751647, AB_2751601, AB_2751797, AB_2751737, AB_2801988, AB_2801967, AB_2811587, AB_2811564, AB_2801869, AB_2733810

References for CD38 Antibody, anti-human, REAfinity™

Publications

  1. 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
  2. Sathish, V. et al. (2014) Inflammation, caveolae and CD38-mediated calcium regulation in human airway smooth muscle. Biochim. Biophys. Acta 1843(2): 346-351
  3. 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

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