Clone:
REA115
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
LY6B

Extended validation for Ly-6B.2 Antibody, anti-mouse, 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 REA115
7/4++
Cells were incubated with an excess of purified unconjugated Ly-6B.2 (REA115) 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 Ly-6B.2. Bone marrow cells from C57BL/6 mice were stained with Ly-6B.2 antibodies and with a suitable counterstaining. As a control, Ly-6B.2 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 Ly-6B.2. Bone marrow cells from C57BL/6 mice were stained with Ly-6B.2 antibodies and with a suitable counterstaining. As a control, Ly-6B.2 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 Ly-6B.2. Bone marrow cells from C57BL/6 mice were stained with Ly-6B.2 antibodies and with a suitable counterstaining. As a control, Ly-6B.2 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 Ly-6B.2. Bone marrow cells from C57BL/6 mice were stained with Ly-6B.2 antibodies and with a suitable counterstaining. As a control, Ly-6B.2 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 Ly-6B.2 (REA115). 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 Ly-6B.2 (REA115). 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 Ly-6B.2 (REA115). 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 Ly-6B.2 Antibody, anti-mouse, REAfinity™

Overview

Clone REA115 recognizes Ly-6B.2, a member of the lymphocyte antigen 6 (Ly-6) superfamily and is located on chromosome 15 as the other members of the murine Ly-6 family. The Ly-6 family members are normally GPI-anchored, cysteine-rich cell surface molecules. Expression of Ly-6B.2 antigen has been reported on neutrophils, bone marrow restricted progenitors, monocytes, and is lost during the differentiation of monoytes into tissue resident macrophages or dendritic cell phenotype. High expression of Ly-6B.2 is also associated with inflammatory cell repertoire including monocytes and certain populations of activated macrophages present during infections.
Additional information: Clone REA115 displays negligible binding to Fc receptors.

Alternative names

LY6B

Detailed product information

Technical specifications

CloneREA115
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenLy-6B.2
Alternative names of antigenLY6B
Distribution of antigenmonocytes, neutrophils
Entrez Gene ID110453
RRIDAB_2652788, AB_2652789, AB_2652792, AB_2652793, AB_2652794, AB_2652795, AB_2652796, AB_2652797, AB_2725908, AB_2652798, AB_2652799

Resources for Ly-6B.2 Antibody, anti-mouse, REAfinity™

Certificates

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

References for Ly-6B.2 Antibody, anti-mouse, REAfinity™

Publications

  1. Rosas, M. et al. (2010) The myeloid 7/4-antigen defines recently generated inflammatory macrophages and is synonymous with Ly-6B. J. Leukoc. Biol. 88: 169-180
  2. Hirsch, S. et al. (1983) Polymorphic expression of a neutrophil differentiation antigen revealed by monoclonal antibody 7/4. Immunogenetics 18(3): 229-239

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