Galectin-3 Expression by Human and Mouse Leucocytes. 1. Surface _ Neutrophils: Human blood was stained with Alexa Fluor™ 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; Dotted Line Histogram) or Alexa Fluor™ 647 Rat Anti-Galectin-3 (Cat. No. 567904; Solid Line Histogram) at 0.25 µg/test. Erythrocytes were lysed with BD FACS™ Lysing Solution. 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 neutrophils. 2. Surface + Intracellular _ PBMC: 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 stained with PE Mouse Anti-Human CD11b and either Alexa Fluor™ 647 Rat IgG2a, κ Isotype Control or Alexa Fluor™ 647 Rat Anti-Galectin-3. A bivariate plot displaying the correlated expression of total cellular Galectin-3 (or Ig Isotype control staining) versus CD11b by intact PBMCs is shown, 3. Surface _ PEC: Thioglycolate-elicited C57BL/6 mouse peritoneal exudate cells (PEC) were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) and stained with either Alexa Fluor™ 647 Rat IgG2a, κ Isotype Control (Dotted Line Histogram) or Alexa Fluor™ 647 Rat Anti-Galectin-3 (Solid Line Histogram) at 0.06 µg/test. DAPI Solution (Cat. No. 564907) was added right before analysis. Surface Galectin-3 expression (or Ig Isotype control staining) on viable (DAPI-negative) PEC is shown. 4. Surface + Intracellular _ PEC: Thioglycolate-elicited PEC were similarly fixed, permeabilized and stained (without DAPI). Total cellular Galectin-3 expression (or Ig Isotype control staining) by intact PEC is shown. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Immunohistofluorescence analysis of Galectin-3 expression in mouse intestine. Following antigen retrieval with BD Pharmingen™ Retrievagen A Buffer (Cat. No. 550524), a formalin-fixed, paraffin-embedded BALB/c mouse intestine section was stained with Alexa Fluor™ 647 Rat Anti-Galectin-3 antibody (Cat. No. 567904; pseudo-colored red; Left Image). The section was counterstained with DAPI (Cat. No. 564907; pseudo-colored blue; Middle Image) as a nuclear stain. The images were captured and merged (Right Image) using a standard epifluorescence microscope system equipped with a 20× objective and software.
Galectin-3 Expression by Human and Mouse Leucocytes. 1. Surface _ Neutrophils: Human blood was stained with Alexa Fluor™ 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690; Dotted Line Histogram) or Alexa Fluor™ 647 Rat Anti-Galectin-3 (Cat. No. 567904; Solid Line Histogram) at 0.25 µg/test. Erythrocytes were lysed with BD FACS™ Lysing Solution. 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 neutrophils. 2. Surface + Intracellular _ PBMC: 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 stained with PE Mouse Anti-Human CD11b and either Alexa Fluor™ 647 Rat IgG2a, κ Isotype Control or Alexa Fluor™ 647 Rat Anti-Galectin-3. A bivariate plot displaying the correlated expression of total cellular Galectin-3 (or Ig Isotype control staining) versus CD11b by intact PBMCs is shown, 3. Surface _ PEC: Thioglycolate-elicited C57BL/6 mouse peritoneal exudate cells (PEC) were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) and stained with either Alexa Fluor™ 647 Rat IgG2a, κ Isotype Control (Dotted Line Histogram) or Alexa Fluor™ 647 Rat Anti-Galectin-3 (Solid Line Histogram) at 0.06 µg/test. DAPI Solution (Cat. No. 564907) was added right before analysis. Surface Galectin-3 expression (or Ig Isotype control staining) on viable (DAPI-negative) PEC is shown. 4. Surface + Intracellular _ PEC: Thioglycolate-elicited PEC were similarly fixed, permeabilized and stained (without DAPI). Total cellular Galectin-3 expression (or Ig Isotype control staining) by intact PEC is shown. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Immunohistofluorescence analysis of Galectin-3 expression in mouse intestine. Following antigen retrieval with BD Pharmingen™ Retrievagen A Buffer (Cat. No. 550524), a formalin-fixed, paraffin-embedded BALB/c mouse intestine section was stained with Alexa Fluor™ 647 Rat Anti-Galectin-3 antibody (Cat. No. 567904; pseudo-colored red; Left Image). The section was counterstained with DAPI (Cat. No. 564907; pseudo-colored blue; Middle Image) as a nuclear stain. The images were captured and merged (Right Image) using a standard epifluorescence microscope system equipped with a 20× objective and software.
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), Immunofluorescence, Immunohistochemistry (Tested During Development)
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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- 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.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
567904 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.
567904 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
567904 Rev.1
Citations & References
View product citations for antibody "567904" on CiteAb
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
567904 Rev. 1
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.