Clone:
REA435
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC, MICS, IHC, IF
Alternative names:
Lgals9, HUAT, LGALS9A, Gal-9, Ecalectin, HOM-HD-21

Extended validation for Galectin-9 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 REA435
9M1-3+
REAL750n/a
Cells were incubated with an excess of purified unconjugated Galectin-9 (REA435) 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 Galectin-9. MOLT-4 cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with Galectin-9 antibodies. As a control, Galectin-9 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 Viobility 405/520 Fixable Dye 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 Galectin-9. MOLT-4 cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with Galectin-9 antibodies. As a control, Galectin-9 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 Viobility 405/520 Fixable Dye 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 Galectin-9. MOLT-4 cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with Galectin-9 antibodies. As a control, Galectin-9 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 Viobility 405/520 Fixable Dye 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 Galectin-9. MOLT-4 cells were first stained with Viobility™ Fixable Dye followed by a suitable counterstaining. Cells were then fixed and permeabilized using the Inside Stain Kit followed by intracellular staining with Galectin-9 antibodies. As a control, Galectin-9 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 Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for Galectin-9 Antibody, anti-human, REAfinity™

Overview

Clone REA435 recognizes the human galectin-9 antigen, a member of the galectin family of carbohydrate-binding proteins, which is also known as ecalectin or tumor antigen HOM-HD-21. The galectin family is characterized by the presence of two or more conserved carbohydrate-recognition domains that bind galactose. Galectin-9 is widely distributed throughout various tissues and circulates at very high levels in the serum. It interacts with various ligands, the best characterized of which is the T cell immunoglobulin mucin 3 (Tim-3) cell surface molecule, widely expressed on innate and adaptive immune cells. Originally described as an eosinophil chemoattractant, Galectin-9 is now known to be an important pleiotropic immune modulator affecting numerous immune cell types. Galectin-9 is thought to be involved in the activation of innate immune responses and the downregulation of Tʜ17 and Tʜ1 responses. It can promote inflammation through triggering proinflammatory cytokine production from monocytes and inducing maturation of macrophages and monocyte-derived dendritic cells. Conversely, Galectin-9 has a major anti-inflammatory role in the induction of apoptosis in activated T cell subsets, as well as in promoting the differentiation of regulatory T cells expressing FoxP3. Galectin-9 is also involved in the apoptosis of tumor cells.
Additional information: Clone REA435 displays negligible binding to Fc receptors.

Alternative names

Lgals9, HUAT, LGALS9A, Gal-9, Ecalectin, HOM-HD-21

Detailed product information

Technical specifications

CloneREA435
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenGalectin-9
Alternative names of antigenLgals9, HUAT, LGALS9A, Gal-9, Ecalectin, HOM-HD-21
Molecular mass of antigen [kDa]40
Entrez Gene ID3965
RRIDAB_2889672, AB_2857692, AB_2857680, AB_2905213, AB_2905212, AB_2889673

Resources for Galectin-9 Antibody, anti-human, REAfinity™

Certificates

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References for Galectin-9 Antibody, anti-human, REAfinity™

Publications

  1. Matsumoto, R. et al. (1998) Human ecalectin, a variant of human galectin-9, is a novel eosinophil chemoattractant produced by T lymphocytes. J. Biol. Chem. 273(27): 16976-16984
  2. Leitner, J. et al. (2013) TIM-3 does not act as a receptor for galectin-9. PLoS Pathog. 9(3): e1003253
  3. Golden-Mason, L. et al. (2013) Galectin-9 functionally impairs natural killer cells in humans and mice. J. Virol. 87(9): 4835-4845

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