50-9116-42

antibody from Invitrogen Antibodies

Targeting: LGALS9 LGALS9A

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

50-9116-42

Provider

Invitrogen Antibodies

Product name

Galectin 9 Monoclonal Antibody (9M1-3), eFluor™ 660, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: This 9M1-3 monoclonal antibody reacts with human Galectin-9. Galectin-9 is one of fifteen members of the galectin family of glycan-binding proteins. It is expressed on activated T helper cells and Foxp3+ T regulatory cells and plays a critical role in the regulation of immune function. Interaction of Galectin-9 with its ligand, TIM3, induces apoptosis in Th1 and Th17, inhibits Th17 polarization and IFN gamma expression, and drives Treg expansion. Galectin-9 is also thought to contribute to the survival and expansion of some tumors by inhibiting the anti-tumor response and promoting immune evasion. Along with Galectin-1 and Galectin-3, high levels of Galectin-9 expression are associated with poor cancer prognosis, making these potential therapeutic targets. Applications Reported: This 9M1-3 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 9M1-3 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

9M1-3

Vial size

100 Tests

Concentration

5 µL/Test

Storage

4° C, store in dark, DO NOT FREEZE!

References

Submitted references

Galectins and their ligands: negative regulators of anti-tumor immunity.

Cedeno-Laurent F, Dimitroff CJ
Glycoconjugate journal 2012 Dec;29(8-9):619-25

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Galectin 9 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Galectin 9 Monoclonal Antibody (9M1-3), eFluor™ 660, eBioscience™ (Product # 50-9116-42) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight. (Panel a: Red), Nuclei (Panel b:Blue) were stained with Hoechst 33342 (Product # H1399, 1:2000 dilution). F-actin (Panel c: Green) was stained with Alexa Fluor™ 488 Phalloidin (Product # A12379, 1:300 dilution). Panel d represents the merged image showing Nuclear as well as cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7A1110LZR) and externally deconvoluted.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Normal human peripheral blood cells were stimulated for 3 days with Human IL-2 Recombinant Protein (Product # 14-8029-81). The cells were stained with Anti-Human CD3 APC-eFluor® 780 (Product # 47-0038-42) and Mouse IgG1 K Isotype Control eFluor® 660 (Product # 50-4714-82) (left) or Anti-Human Galectin-9 eFluor® 660 (right). Viable cells in the lymphocyte gate, as determined by Fixable Viability Dye eFluor® 506 (Product # 65-0866-14), were used for analysis.