Clone:
REA526
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
Ly-6G.1, Ly6g, Gr-1

Extended validation for Ly-6G 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 REA526
REA810-
1A8++
RB6-8C5-
Cells were incubated with an excess of purified unconjugated Ly-6G (REA526) 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 Anti-Ly-6G. Bone marrow cells from C57BL/6 mice were stained with Anti-Ly-6G antibodies and with a suitable counterstaining. As a control, Anti-Ly-6G 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 Anti-Ly-6G. Bone marrow cells from C57BL/6 mice were stained with Anti-Ly-6G antibodies and with a suitable counterstaining. As a control, Anti-Ly-6G 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 Anti-Ly-6G. Bone marrow cells from C57BL/6 mice were stained with Anti-Ly-6G antibodies and with a suitable counterstaining. As a control, Anti-Ly-6G 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 Anti-Ly-6G. Bone marrow cells from C57BL/6 mice were stained with Anti-Ly-6G antibodies and with a suitable counterstaining. As a control, Anti-Ly-6G 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-6G (REA526). 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-6G (REA526). 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-6G (REA526). 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-6G Antibody, anti-mouse, REAfinity™

Overview

Clone REA526 recognizes the mouse lymphocyte antigen 6G (Ly-6G) antigen, a 21–25 kDa glycosylphosphatidylinositol (GPI)-linked protein which is highly expressed on neutrophilic granulocytes. Expression greatly increases during granulocyte maturation. To a lower extent, Ly-6G is also expressed during the development of monocytes. In mouse bone marrow samples, Ly-6G is primarily detected on granulocytes, but not on lymphocytes or erythrocytes. Ligation of Ly-6G in animals with an underlying background inflammation (induced by TNF-α) leads to profound cardiovascular and respiratory responses, including severe breathing difficulties, leukocyte and platelet aggregation, and coagulation, leading to microvessel obstruction and subsequent death.
Additional information: Clone REA526 displays negligible binding to Fc receptors.

Alternative names

Ly-6G.1, Ly6g, Gr-1

Detailed product information

Technical specifications

CloneREA526
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody, human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Speciesmouse
AntigenLy-6G
Alternative names of antigenLy-6G.1, Ly6g, Gr-1
Molecular mass of antigen [kDa]10
Distribution of antigenmonocytes, macrophages, granulocytes
Entrez Gene ID546644
RRIDAB_2733090, AB_2727967, AB_2727584, AB_2727602, AB_2751964, AB_2751917, AB_2784431, AB_2752201, AB_2752182, AB_2784432, AB_2801997, AB_2811549, AB_2733089

Resources for Ly-6G Antibody, anti-mouse, REAfinity™

Certificates

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References for Ly-6G Antibody, anti-mouse, REAfinity™

Publications

  1. Fleming, T. J. et al. (1993) Characterization of two novel Ly-6 genes. Protein sequence and potential structural similarity to alpha-bungarotoxin and other neurotoxins. J. Immunol. 150(12): 5379-5390
  2. Carr, K. D. et al. (2011) Specific depletion reveals a novel role for neutrophil-mediated protection in the liver during Listeria monocytogenes infection. Eur. J. Immunol. 41(9): 2666-2676
  3. Tanaka, Y. et al. (2012) Stimulation of Ly-6G on neutrophils in LPS-primed mice induces platelet-activating factor (PAF)-mediated anaphylaxis-like shock. J. Leukoc. Biol. 91(3): 485-494

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