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LGALS3 Antibody (OASA07770)

  • Catalog#: OASA07770
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Gene Symbol:
LGALS3
NCBI Gene Id:
3958
Protein Name:
Galectin-3
Swissprot Id:
P17931
Protein Accession #:
NP_002297.2
Alias Symbols:
L31, GAL3, MAC2, CBP35, GALBP, GALIG, LGALS2, LGALS3
Description of Target:
RABBIT ANTI HUMAN GALECTIN-3
Protein Size (# AA):
250
Host:
Rabbit
Clonality:
Polyclonal
Application:
ELISA, WB
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express LGALS3.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express LGALS3.
Immunogen:
The immunogen for anti-LGALS3 antibody: recombinant human galectin-3
Predicted Species Reactivity:
Human
Predicted Homology Based on Immunogen Sequence:
Human
Product Format:
Purified IgG - lyophilised
Reconstitution and Storage:
Reconstitution: Reconstitute with 0.1ml distilled water. For long term storage the addition of 0.09% sodium azide is recommended.
Storage: Prior to reconstitution store at +4oC.
After reconstitution store at -20oC.
Storage in frost-free freezers is not recommended. This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Datasheets/Manuals:
Printable datasheet for anti-LGALS3 antibody - OASA07770
Specificity:
GALECTIN-3
Isotype:
Polyclonal IgG
Application Info:
ELISA : This product may be used in an indirect ELISA or as the capture reagent in a sandwich ELISA with This product B as the detection antibody and OPSA11036 as the standard.
Western Blot: This product may be used under either reducing or non-reducing conditions with OPSA11036 as the positive control.
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Preservative Stabilisers: None present
Antiserum Preparation: Antiserum to human galectin-3 was raised by repeated immunisation of rabbits with highly purified antigen. Purified IgG was prepared from whole serum by affinity chromatography.
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Approx Protein Conc: IgG concentration 1.0mg/ml
Buffer Solutions: Phosphate buffered saline pH7.2
Protocol Information:
Citation: 1: Kiefer FW, Zeyda M, Gollinger K, Pfau B, Neuhofer A, Weichhart T, Säemann MD, Geyeregger R, Schlederer M, Kenner L, Stulnig TM. Neutralization of osteopontininhibits obesity-induced inflammation and insulin resistance. Diabetes. 2010Apr;59(4):935-46. Epub 2010 Jan 27. PubMed PMID: 20107108; PubMed Central PMCID: PMC2844841.
Species: Mouse
Experiment Name: Immunoflourescence, immunohistochemistry, tunel staining, and flow cytometry.
Experiment Background: 1. The number of macrophages in GWAT, as determined by immunofluorescence (F4/80+) and immunohistochemistry (Mac-2) 2. Apoptotic cells were determined by tunel staining (green), macrophages were stained red by immunoflourescence using anti-F4/80 monoclonal antibody on frozen sections.3. Macrophages were stained red by immunoflourescence using Mac-2 monoclonal antibody on frozen sections of livers isolated from high-fat diet–fed mice after anti-OPN or control treatment. Apoptotic cells were determined by TUNEL staining (green).4. Obese high-fat diet–fed (HF) mice were treated with OPN-neutralizing (Anti-OPN) or control (n = 8 per group) antibody. STAT phosphorylation was determined by immunhistochemical analysis of pSTAT(Tyr705) on paraffin sections of livers isolated from high-fat diet–fed mice after anti-OPN or control reatment.
Experimental Steps: Frozen sections were prepared from murine GWAT and liver. Sections were stained with rat anti-mouse F4/80 and Mac-2 IgG antibodies (Oxford, U.K. and Cedarlane, Burlington, ON, Canada, respectively). Primary antibodies were detected with AlexaFluor 488 or AlexaFluor 594 goat anti-rat IgG antibodies (Molecular Probes, Eugene, OR). As a negative control, isotype control staining was done on selected sections. Nuclei were visualized by DAPI staining. Slides were mounted in Vectashield (Vector Laboratories, Burlingame, CA) and examined under a fluorescence microscope (Leica, Wetzlar, Germany). Macrophage infiltration in adipose tissue and liver was quantified by calculating the ratio of F4/80- and Mac-2–positive cells to totalnuclei. Apoptotic cells were stained on frozen sections using the Fluorescin In Situ Cell Dection Kit from Roche, according to manufacturer’s instructions, in parallel with double staining for F4/80 and Mac-2, respectively.For paraffin sections, GWAT and liver samples were fixed with neutral buffered 4% paraformaldehyde and were paraffin-embedded. Hematoxylin and eosin staining was performed in liver. After dewaxation and rehydration, immunohistochemical staining for Mac-2 and pSTAT3 (Tyr 705) (Cell Signaling, Danvers, MA) was performed on adipose tissue and liver sections, respectively, using the ABC kit (Vector Laboratories) according to the manufacturer’s recommendations. As a negative control, staining was performed on selected sections with isotype control. Samples were analyzed with standard light microscopy and a Zeiss Axio-Imager Z1 microscope system with a charge-coupled device camera and a TissueFAXS automated acquisition system (TissueGnostics, Vienna, Austria), respectively. The percentage of pSTAT3-positive cells of 1,000 cells was determined using HistoQuest software (TissueGnostics).Stromal vascular cells (SVCs) of GWAT were isolated by collagenase digestion and centrifugation to remove adipocytes. The percentage of macrophages within SVCs was determined using phycoerythrin-conjugated anti-F4/80 mAb (Abd ) and standard flow cytometric procedures.
Number Of Protocols: 1
Target Reference:
1. Dumic, J. et al. (2006) Galectin-3: an open-ended story. Biochim. Biophys. Acta. 1760: 616-635.

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