深圳市睿盟创新生物科技有限公司

【 Galectin-3 Blocking Antibody 8-1 】

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产品编号:

XS-001-Ab05-C-100

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Specification
Product name	Galectin-3 Blocking Antibody 8-1
Description	Mouse monoclonal antibody raised against a full-length recombinant Galectin-3. Monoclonal antibodies block the binding of Galectin-3 to laminin.
Immunogen	Recombinant full length protein. This information is proprietary to RMi and/or its suppliers.
Source/Isotype	Mouse IgG2b
Reactivity	Human
Specificity	Detects human Galectin-3 in ELISAs. In indirect ELISAs, no cross-reactivity with recombinant human Galectin-1, Galectin-9 is observed.
Purification	Protein G purified from hybridoma culture supernatant
Application	ELISA, others unspecified.
Form	Liquid
Storage Buffer	In 1✕PBS, pH 7.2
Storage Instruction	Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or lower long term.. Aliquot to avoid repeated freezing and thawing.
Size	25ug, 100ug, 500ug

Data Background
The galectins constitute a large family of carbohydrate-binding proteins with specificity for N-acetyl-lactosamine-containing glycoproteins. At least 14 mammalian galectins, which share structural similarities in their carbohydrate recognition domains (CRD), have been identified. The galectins have been classified into the prototype galectins (-1, -2, -5, -7, -10, -11, -13, -14), which contain one CRD and exist either as a monomer or a noncovalent homodimer; the chimera galectins (Galectin-3) containing one CRD linked to a nonlectin domain; and the tandem-repeat galectins (-4, -6, -8, -9, -12) consisting of two CRDs joined by a linker peptide. Members of the galectin family have been implicated in a variety of biological functions including cell adhesion, growth regulation, cytokine production, T-cell apoptosis, and immune responses. Galectin-3 is involved in several diverse biological functions. Galectin-3 binds IgE. Galectin-3 is an unusual protein in that it can be found both extracellularly and intracellularly. Intracellularly, galectin-3 can localize to the cytoplasm, nucleus, or both, depending on cell type and experimental conditions. Nuclear galectin-3 has been identified as a pre-mRNA splicing factor. Galectin-3 production has been shown to increase during inflammation and in obesity, and the protein itself can have an inflammatory effect under certain conditions. Galectin-3 forms a complex with α3, β1 integrin to act as a surface receptor on endothelial cells for the NG2 proteoglycan, which triggers cell motility and angiogenesis. In addition to these functions, galectin-3 is also a required factor for the terminal differentiation of epithelial cells.

Supporting Data

1. Indirect ELISA protocol

1.1          Reagents：
1.1.1     Coating Buffer: Carbonate Buffer (2.93 g NaHCO₃, 1.59 g Na₂CO₃, 1L ddH₂O)
1.1.2     Diluent: PBS (137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5mM KH₂PO₄)
1.1.3     Blocking Solution: 1% BSA in PBS
1.1.4     Secondary Ab: anti mouse IgG-HRP(Sigma A2554-1ML)
1.2          Assay Protocol：
Note: The following protocol is a guideline, user need to determine their optimal experimental condition for best performance.
1.2.1     Coat plate with Recombinant protein 0.2ug/ml, 100ul/well, incubate at 4℃ for overnight.
1.2.2     Wash the plate with PBST (0.05% Tween-20) once.
1.2.3     Add 200μL of blocking buffer each well, incubate the plate at 37℃ for 2hr or 4℃ overnight.
1.2.4     Add sample 100ul/well, incubate the plate at 37℃ for 1hr.
1.2.5     Wash the plate with PBST (0.05% Tween-20) 3 times.
1.2.6     Add secondary Ab, 1:10k diluted anti-mouse IgG-HRP, 100ul/well, incubate the plate at 37℃ for 0.5hr.
1.2.7     Wash the plate with PBST (0.05% Tween-20) 3 times.
  1.2.8     Add 100ul/well TMB, incubate at room temperature for 10min, add 50ul/well 1N HCl or 2% H₂SO₄.
1.2.9     Read absorbance at 450nm.
2. Galectin-3 Antibody binding/blocking assay protocol
2.1 Reagents：
2.1.1 Coating Buffer: PBS (137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5mM KH2PO4)
2.1.2 Laminin: Corning, 354232
2.1.3 Diluent: 1% BSA in PBS
2.1.4 Blocking Solution: 1% BSA in PBS
2.1.5 Secondary Ab: anti mouse IgG-HRP(Sigma A2554-1ML)
   2.2Assay Protocol
Note: The following protocol is a guideline, user need to determine their optimal experimental condition for
best performance
2.2.1 Coat plate with Laminin 5ug/ml, 100ul/well, Incubate at 4℃ for overnight.
2.2.2 Wash the plate with PBST (0.05% Tween-20) once.
2.2.3 Add 200μL of blocking buffer each well, incubate the plate at 37℃ for 2hr or 4℃ overnight.
2.2.4 Add the plate with a series of anti-Galectin-3-Ab 50ul/well, and then add 2.0ug/ml Galectin-3 solution
50ul/well, incubated at 37℃ for 1hr.
2.2.5 Wash the plate with PBST (0.05% Tween-20) 3 times
2.2.6 Add secondary Ab, 1:10k diluted anti-mouse IgG-HRP, 100ul/well, incubate the plate at 37℃ for 0.5hr.
2.2.7 Wash the plate with PBST (0.05% Tween-20) 3 times
2.2.8 Add 100ul/well TMB, incubate at room temperature for 10min, add 50ul/well 1N HCl or 2% H2SO4.
2.2.9 Read absorbance at 450nm.

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