Clone:
REA431
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IHC, IF
Alternative names:
MICL, CLL-1, DCAL-2

Extended validation for CD371 (CLEC12A) 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 REA431
687317-
50C1++
HB3-
Cells were incubated with an excess of purified unconjugated CD371 (CLEC12A) (REA431) 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.

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 CD371 (CLEC12A). Human peripheral blood mononuclear cells (PBMCs) were stained with CD371 (CLEC12A) antibodies and with a suitable counterstaining. As a control, CD371 (CLEC12A) 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 propidium iodide 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 CD371 (CLEC12A). Human peripheral blood mononuclear cells (PBMCs) were stained with CD371 (CLEC12A) antibodies and with a suitable counterstaining. As a control, CD371 (CLEC12A) 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 propidium iodide 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 CD371 (CLEC12A). Human peripheral blood mononuclear cells (PBMCs) were stained with CD371 (CLEC12A) antibodies and with a suitable counterstaining. As a control, CD371 (CLEC12A) 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 propidium iodide 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 CD371 (CLEC12A). Human peripheral blood mononuclear cells (PBMCs) were stained with CD371 (CLEC12A) antibodies and with a suitable counterstaining. As a control, CD371 (CLEC12A) 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 propidium iodide 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 CD371 (CLEC12A) (REA431). 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 CD371 (CLEC12A) (REA431). 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 CD371 (CLEC12A) (REA431). 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 CD371 (CLEC12A) Antibody, anti-human, REAfinity™

Overview

Clone REA431 recognizes CD371, the human C-type lectin domain family 12 member A (CLEC12A) antigen, a single-pass type II membrane protein which is also known as C-type lectin-like molecule 1 (CLL-1), dendritic cell-associated lectin 2 (DCAL2), or myeloid inhibitory C-type lectin-like receptor (MICL). CD371 (CLEC12A) is a immunoreceptor tyrosine-based inhibitory motif (ITIM) containing C-type lectin receptor encoded within the dectin-1 cluster. This receptor binds unknown endogenous ligands and is widely expressed on innate immune cells, including neutrophils, monocytes, macrophages, and dendritic cells (DCs). There are conflicting data on its expression on human natural killer (NK) cells. Its intracellular ITIM sequence recruits the tyrosine phosphatases SHP-1 and SHP-2 to negatively regulate Syk signaling. Because the expression of CD371 (CLEC12A) can be downregulated under inflammatory conditions, this receptor might be involved in the control of myeloid cell activation. CD371 (CLEC12A) has been shown to enhance LPS responses and to induce expression of CCR7, a chemokine involved in DC migration. It has been proposed that signals from CD371 (CLEC12A) may synergize with signals from other receptors to facilitate DC responses such as migration and T cell stimulation.
Additional information: Clone REA431 displays negligible binding to Fc receptors.

Alternative names

MICL, CLL-1, DCAL-2

Detailed product information

Technical specifications

CloneREA431
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenCD371 (CLEC12A)
Alternative names of antigenMICL, CLL-1, DCAL-2
Molecular mass of antigen [kDa]31
Distribution of antigendendritic cells, macrophages, monocytes, neutrophils
Entrez Gene ID160364
RRIDAB_2801968, AB_2876960, AB_2876961, AB_2819704, AB_2819696, AB_2657807, AB_2657808, AB_2657809, AB_2657810, AB_2801989

References for CD371 (CLEC12A) Antibody, anti-human, REAfinity™

Publications

  1. Lahoud, M. H. et al. (2009) The C-type lectin Clec12A present on mouse and human dendritic cells can J. Immunol. 182(12): 7587-7594
  2. Neumann, K. et al. (2014) Clec12a is an inhibitory receptor for uric acid crystals that regulates inflammation in response to cell death. Immunity 40(3): 389-399
  3. Bakker, A. B. et al. (2004) C-type lectin-like molecule-1: a novel myeloid cell surface marker associated with acute myeloid leukemia. Cancer Res. 64(22): 8443-8450

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