Dynabeads Protein G (10 µL, Cat# 10007D, Thermo) was incubated with 0.2 μg of LCN2 mAb (Cat# MAB18571, R&D) in 100 µL PBS for 60 min at room temperature [57 (link)]. For the competitive interaction between LCN2 mAb and mouse LCN2 protein, LCN2 mAb bound on the Dynabeads was incubated with 0.06 μg of recombinant mouse LCN2 protein (Cat# 1857-LC, R&D) and mouse LCN2 mAb epitope peptide (YNVTSILVRDQDQGCRYWIRT, synthesized by AnaSpec, Fremont, CA, USA) in 10X, 100X, or 1000X molar ratio of LCN2 protein in 100 µL PBST for 16 h at 4 °C. For the competitive interaction between LCN2 mAb and human LCN2 protein, recombinant human LCN2 protein (Cat# 1757-LC, R&D) and human LCN2 mAb epitope peptide (YNVTSVLFRKKKCDYWIRT, synthesized by AnaSpec) were used. The immunoprecipitated mouse or human LCN2 protein was detected by Western blot using goat anti-mouse LCN2 antibody (Cat# AF1857, R&D; 1:1000 dilution in Western Antibody Dilution Buffer) or goat anti-human LCN2 antibody (Cat# AF1757, R&D; 1:1000 dilution in Western Antibody Dilution Buffer), and an EasyBlot anti-goat IgG kit (Cat# GTX228910-01, GeneTex).
Goat anti human lcn2 antibody
Manufactured by R&D Systems
Sourced in United States
Goat anti-human LCN2 antibody is a primary antibody that specifically binds to the human lipocalin-2 (LCN2) protein. LCN2 is a secreted glycoprotein involved in various biological processes. This antibody can be used for the detection and quantification of LCN2 in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse lcn2 antibody, by R&D Systems (1 mentions)
Easyblot anti goat igg kit, by GeneTex (1 mentions)
Dynabeads protein g, by Thermo Fisher Scientific (1 mentions)
Af1757, by R&D Systems (1 mentions)
Vectastain elite abc kit, by Vector Laboratories (1 mentions)
Mas coated slides, by Matsunami (1 mentions)
Immobilon, by Merck Group (1 mentions)
Mouse anti β actin antibody, by Merck Group (1 mentions)
Microcon ym 3, by Merck Group (1 mentions)
Ecl plus western blotting detection reagent, by GE Healthcare (1 mentions)
Block ace, by Sumitomo Pharma (1 mentions)
3 protocols using goat anti human lcn2 antibody
1
Competitive Binding Assay for LCN2 Antibody
2
Immunohistochemical Analysis of Endometrial and Fetal Membrane Tissues
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Endometrial tissues that had been fixed in 4% paraformaldehyde and embedded in paraffin were sectioned at 6 μm, and mounted onto MAS-coated slides (Matsunami Glass Ind, Osaka, Japan) [28 (link)].
Frozen fetal membranes were also sectioned at 6 μm in cryostat, mounted on glass slides, and then fixed in 4% paraformaldehyde. Antigen retrieval was initially performed in a heat-treated (95°C for 5 min) sodium citrate buffer
(0.01 M, pH 6.0). The sections were treated with 3% H2O2/methanol at room temperature for 30 min to reduce endogenous peroxidase activity and then in Block Ace at room temperature for 1 h to quench
non-specific staining. The sections were incubated with goat anti-human LCN2 antibody (4 μg/ml, AF1757, R&D Systems), goat anti-human SLC22A17 antibody (4 μg/ml, OAEB00639, Aviva Systems Biology, San Diego, CA, USA) or normal
goat IgG (4 μg/ml) at 4°C for 12 h. After the primary antibody incubation, the slides were incubated with biotin-conjugated donkey anti-goat IgG (GE Healthcare UK Ltd.) at room temperature for 1 h. Specific signals were visualized
using the VECTASTAIN Elite ABC Kit (Vector Laboratories, Burlingame, CA, USA). The tissue sections were counterstained with hematoxylin.
Frozen fetal membranes were also sectioned at 6 μm in cryostat, mounted on glass slides, and then fixed in 4% paraformaldehyde. Antigen retrieval was initially performed in a heat-treated (95°C for 5 min) sodium citrate buffer
(0.01 M, pH 6.0). The sections were treated with 3% H2O2/methanol at room temperature for 30 min to reduce endogenous peroxidase activity and then in Block Ace at room temperature for 1 h to quench
non-specific staining. The sections were incubated with goat anti-human LCN2 antibody (4 μg/ml, AF1757, R&D Systems), goat anti-human SLC22A17 antibody (4 μg/ml, OAEB00639, Aviva Systems Biology, San Diego, CA, USA) or normal
goat IgG (4 μg/ml) at 4°C for 12 h. After the primary antibody incubation, the slides were incubated with biotin-conjugated donkey anti-goat IgG (GE Healthcare UK Ltd.) at room temperature for 1 h. Specific signals were visualized
using the VECTASTAIN Elite ABC Kit (Vector Laboratories, Burlingame, CA, USA). The tissue sections were counterstained with hematoxylin.
3
Endometrial Lysate and Luminal Fluid Protein Analysis
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Protein samples (20 μg) from endometrial lysates in reduced condition and centrifugally concentrated (Microcon YM-3; Millipore, Bedford, MA, USA) proteins from luminal fluid (4 μg) in reduced or non-reduced condition were
separated on 12.5% SDS-PAGE and transferred onto nitrocellulose membrane (Immobilon; Millipore) [1 (link), 25 (link), 28 (link)]. After
blocking with Block Ace (Dainippon Sumitomo Pharma) at room temperature for 1 h, the membranes were incubated with goat anti-human LCN2 antibody (200 ng/ml; AF1757, R&D systems, Minneapolis, MN, USA) or mouse anti-β-actin
antibody (30 ng/ml; Sigma-Aldrich, St. Louis, MO, USA) at 4°C for 12 h. After incubation, the membrane was washed three times in TBS-Tween 20 and incubated with horseradish peroxidase-conjugated donkey anti-goat IgG or goat
anti-mouse IgG (Amersham Biosciences, Piscataway, NJ, USA) at room temperature for 1 h. Signals were detected using ECL Plus Western Blotting Detection Reagents (GE Healthcare UK, Buckinghamshire, UK).
separated on 12.5% SDS-PAGE and transferred onto nitrocellulose membrane (Immobilon; Millipore) [1 (link), 25 (link), 28 (link)]. After
blocking with Block Ace (Dainippon Sumitomo Pharma) at room temperature for 1 h, the membranes were incubated with goat anti-human LCN2 antibody (200 ng/ml; AF1757, R&D systems, Minneapolis, MN, USA) or mouse anti-β-actin
antibody (30 ng/ml; Sigma-Aldrich, St. Louis, MO, USA) at 4°C for 12 h. After incubation, the membrane was washed three times in TBS-Tween 20 and incubated with horseradish peroxidase-conjugated donkey anti-goat IgG or goat
anti-mouse IgG (Amersham Biosciences, Piscataway, NJ, USA) at room temperature for 1 h. Signals were detected using ECL Plus Western Blotting Detection Reagents (GE Healthcare UK, Buckinghamshire, UK).
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