Galectin-3 Expression by Human and Mouse Leucocytes 1. Surface_Neutrophils Human blood was stained with Alexa Fluor™ 488 Rat IgG2a, κ Isotype Control (Cat. No. 557676; dashed histogram) or Alexa Fluor™ 488 Rat Anti-Galectin-3 (Cat. No. 568406/568407; solid line histogram) at 0.125 µg/test. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A histogram showing Surface Galectin-3 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable cells. 2. Surface + Intracellular_PBMC Peripheral blood mononuclear cells (PBMC) were preincubated with Human BD Fc Block™ (Cat. No. 564219/564220), fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). Cells were then stained with PE Mouse Anti-Human CD11b (Cat. No. 555388) and with either Alexa Fluor™ 488 Rat IgG2a, κ Isotype Control or Alexa Fluor™ 488 Rat Anti-Galectin-3. The bivariate pseudocolor density plot shows the correlated expression of total cellular Galectin-3 (or Ig Isotype control staining) versus CD11b by gated intact PBMC. 3. Surface_PEC Thioglycolate-elicited C57BL/6 Mouse peritoneal exudate cells (Thio-PEC) were preincubated with Purified Rat Anti-Mouse CD16/CD32 Ab (Mouse BD Fc Block™) and stained with either Alexa Fluor™ 488 Rat IgG2a, κ Isotype Control (dashed line histogram) or Alexa Fluor™ 488 Rat Anti-Galectin-3 (solid line histogram) at 0.125 µg/test. The histogram shows Surface Galectin-3 expression (or Ig Isotype control staining) on viable (DAPI-negative) [Cat. 564907] Thio-PEC. 4. Surface + Intracellular_PEC Thio-PEC were similarly fixed, permeabilized and stained (without DAPI). The histogram shows total cellular Galectin-3 expression (or Ig Isotype control staining) by intact Thio-PEC. Flow cytometry and analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown are not lot specific.
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Galectin-3 Expression by Human and Mouse Leucocytes 1. Surface_Neutrophils Human blood was stained with Alexa Fluor™ 488 Rat IgG2a, κ Isotype Control (Cat. No. 557676; dashed histogram) or Alexa Fluor™ 488 Rat Anti-Galectin-3 (Cat. No. 568406/568407; solid line histogram) at 0.125 µg/test. Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A histogram showing Surface Galectin-3 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable cells. 2. Surface + Intracellular_PBMC Peripheral blood mononuclear cells (PBMC) were preincubated with Human BD Fc Block™ (Cat. No. 564219/564220), fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). Cells were then stained with PE Mouse Anti-Human CD11b (Cat. No. 555388) and with either Alexa Fluor™ 488 Rat IgG2a, κ Isotype Control or Alexa Fluor™ 488 Rat Anti-Galectin-3. The bivariate pseudocolor density plot shows the correlated expression of total cellular Galectin-3 (or Ig Isotype control staining) versus CD11b by gated intact PBMC. 3. Surface_PEC Thioglycolate-elicited C57BL/6 Mouse peritoneal exudate cells (Thio-PEC) were preincubated with Purified Rat Anti-Mouse CD16/CD32 Ab (Mouse BD Fc Block™) and stained with either Alexa Fluor™ 488 Rat IgG2a, κ Isotype Control (dashed line histogram) or Alexa Fluor™ 488 Rat Anti-Galectin-3 (solid line histogram) at 0.125 µg/test. The histogram shows Surface Galectin-3 expression (or Ig Isotype control staining) on viable (DAPI-negative) [Cat. 564907] Thio-PEC. 4. Surface + Intracellular_PEC Thio-PEC were similarly fixed, permeabilized and stained (without DAPI). The histogram shows total cellular Galectin-3 expression (or Ig Isotype control staining) by intact Thio-PEC. Flow cytometry and analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown are not lot specific.
Product Details
BD Pharmingen™
CBP 35; LGALS3; Lgals3; MAC2; Mac-2; gal-3; gal3
Human (QC Testing), Mouse (Tested in Development)
Rat IgG2a, κ
Mouse Galectin-3
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Alexa Fluor® 488 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557676
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Human BD Fc Block™ RUO
Size
50 µg
Cat No.
564219
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
PE Mouse Anti-Human CD11b RUO
Size
100 Tests
Cat No.
555388
568406 Rev. 1
Antibody Details
M3/38
The M3/38 monoclonal antibody specifically recognizes Galectin-3 (Gal-3 or gal3) which is also known as Galactose-specific lectin 3, Mac-2, MAC2, and Carbohydrate-binding protein 35 (CBP 35). Galectin-3 is an ~30-35 kDa protein that includes an N-terminal proline-rich tandem repeat domain as well as a C-terminal region with one carbohydrate recognition domain (CRDs). Galectin-3 is encoded by Lgals3 (lectin, galactose binding, soluble 3) which belongs to the galectin family. It is expressed by a variety of cells including activated macrophages, dendritic cells (DCs), granulocytes, mast cells, and epithelial cells. Galectin-3 can be detected in monomeric or multimeric forms in cytoplasmic, nuclear, cell surface and extracellular locations. Galectin-3 is a β-galactoside-binding lectin that has multiple intracellular and extracellular functions implicated in biological processes including RNA splicing as well as cellular chemoattraction and adhesion, activation, growth, proliferation, differentiation, and apoptosis. Upregulated Galectin-3 expression is implicated in inflammation and disease states including cancer progression and metastasis.
568406 Rev. 1
Format Details
Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
568406 Rev.1
Citations & References
Development References (8)
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Demotte N, Wieërs G, Van Der Smissen P, et al. A galectin-3 ligand corrects the impaired function of human CD4 and CD8 tumor-infiltrating lymphocytes and favors tumor rejection in mice.. Cancer Res. 2010; 70(19):7476-88. (Clone-specific: Flow cytometry). View Reference
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Dong R, Zhang M, Hu Q, et al. Galectin-3 as a novel biomarker for disease diagnosis and a target for therapy (Review).. Int J Mol Med. 2018; 41(2):599-614. (Biology). View Reference
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Flotte TJ, Springer TA, Thorbecke GJ. Dendritic cell and macrophage staining by monoclonal antibodies in tissue sections and epidermal sheets. Am J Pathol. 1983; 111(1):112-124. (Clone-specific: Immunohistochemistry). View Reference
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Gray RM, Davis MJ, Ruby KM, Voss PG, Patterson RJ, Wang JL. Distinct effects on splicing of two monoclonal antibodies directed against the amino-terminal domain of galectin-3.. Arch Biochem Biophys. 2008; 475(2):100-8. (Clone-specific: Functional assay, Western blot). View Reference
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Ho MK, Springer TA. Mac-2, a novel 32,000 Mr mouse macrophage subpopulation-specific antigen defined by monoclonal antibodies.. J Immunol. 1982; 128(3):1221-8. (Immunogen: Flow cytometry, Immunoprecipitation, Radioimmunoassay). View Reference
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Johannes L, Jacob R, Leffler H. Galectins at a glance.. J Cell Sci. 2018; 131(9):jcs208884. (Biology). View Reference
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Karlsson A, Follin P, Leffler H, Dahlgren C. Galectin-3 activates the NADPH-oxidase in exudated but not peripheral blood neutrophils.. Blood. 1998; 91(9):3430-8. (Clone-specific: Flow cytometry). View Reference
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Newlaczyl AU, Yu LG. Galectin-3--a jack-of-all-trades in cancer.. Cancer Lett. 2011; 313(2):123-8. (Biology). View Reference
568406 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
