The following commercial materials were used: Alexa Fluor 546 conjugated anti-rabbit IgG antibody and Alexa Fluor 488-conjugated anti-goat IgG antibodies were from Thermo Fisher Scientific (Yokohama, Japan); anti-mast cell tryptase (V-13) antibody against mMCP-6 was from Santa Cruz Biotechnology (Santa Cruz, CA); anti-PAR2 antibody was from Bioss Inc. (Woburn, MA); anti-NF-H antibody was from Gene Tex Inc. (Irvine, CA); anti-GFP antibody was from Novus Biologicals (Centennial, CO); PAR2 antagonist peptide (FSLLRY-NH2) (TOCRIS), PAR2 agonist peptide (SLIGKV-NH2) (TOCRIS) and ATP Detection Assay Kit (Cayman Chemicals) were from Funakoshi Co., Ltd. (Tokyo, Japan); WEHI-3 cells were from American Type Culture Collection; RNA extraction kit (Thermo Fisher Scientific) and real time PCR kit (TAKARA) were from Takara Bio Inc. (Kusatsu, Japan); tryptase substrate (S-2288) was from Chromogenix (Milano, Italy); ATP, sodium monoiodoacetate (MIA) and apyrase were from Sigma-Aldrich Japan (Tokyo, Japan).
Anti gfp antibody
Manufactured by Novus Biologicals
Sourced in United States
The Anti-GFP antibody is a laboratory reagent used for detecting and visualizing green fluorescent protein (GFP) in biological samples. It is a highly specific antibody that binds to GFP, allowing researchers to track and monitor the expression and localization of GFP-tagged proteins in cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Adenosine triphosphate (atp), by Merck Group (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Ecl reagent, by Merck Group (2 mentions)
Real time pcr kit, by Takara Bio (1 mentions)
Apyrase, by Merck Group (1 mentions)
Alexa fluor 546 conjugated anti rabbit igg antibody, by Thermo Fisher Scientific (1 mentions)
Wehi 3 cells, by American Type Culture Collection (1 mentions)
Atp detection assay kit, by Cayman Chemical (1 mentions)
Sodium monoiodoacetate mia, by Merck Group (1 mentions)
Anti mouse caspase 1, by Adipogen (1 mentions)
Anti asc antibody, by Santa Cruz Biotechnology (1 mentions)
Anti f4 80 antibody, by Abcam (1 mentions)
Tamoxifen, by Merck Group (1 mentions)
4 hydroxytamoxifen, by Merck Group (1 mentions)
Mcc950, by Merck Group (1 mentions)
Anti mouse il 1β antibody, by R&D Systems (1 mentions)
Cyanine3 (cy3), by Jackson ImmunoResearch (1 mentions)
Anti nlrp3, by Adipogen (1 mentions)
Anti kiss1, by Abcam (1 mentions)
Csu 22, by Nikon (1 mentions)
6 protocols using anti gfp antibody
1
Mast Cell Tryptase and PAR2 Signaling
2
NLRP3 Inflammasome Activation Assay
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LPS (L4391 for cell culture, L3012 for mice), ATP, MCC950, tamoxifen, and 4-hydroxytamoxifen were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-F4/80 antibody (Cat# ab6640, RRID:AB_1140040) was obtained from Abcam (Cambridge, United Kingdom). Cy3- (Cat# 712-165-050, RRID:AB_2340666) and Alexa Fluor 488-conjugated (Cat# 712-545-150, RRID:AB_2340683) anti-rat IgG antibodies were purchased from Jackson ImmunoResearch (West Grove, PA, USA). FITC-dextran (46944) and TRITC-dextran (T1037) were obtained from Sigma-Aldrich. The IL-1 receptor antagonist (Cat# CYT-203) was purchased from ProSpec (Rehovot, Israel). Anti-mouse caspase-1 (Cat# AG-20B-0042, RRID:AB_2490248) and anti-NLRP3 (Cat# AG-20B-0014, RRID:AB_2490202) antibodies were obtained from AdipoGen Life Sciences (San Diego, CA, USA). Anti-mouse IL-1β antibody (Cat# AF-401-NA, RRID:AB_416684) was obtained from R&D Systems (Minneapolis, MN, USA), and anti-ASC antibody (Cat# SC-22514-R, RRID:AB_2174874) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-GFP antibody (Cat# NB600-308, RRID:AB _10003058) was obtained from NOVUS Biologicals.
3
Western Blot Analysis of GFP Expression
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Ten micrograms of protein for each sample was resolved on a 15% SDS-PAGE, transferred onto a PVDF membrane (Millipore, Billerica, MA) and blocked with 5% w/v skimmed milk protein prepared in 0.1% TBST (room temperature, 1 hr). Subsequently, the membrane was incubated overnight with anti-GFP antibody (Novus, USA—Cat. No. NB100-2220; 1:1000 dilution in 0.1% TBST) at 4°C on a rocker-shaker. Following three washes of 5 minutes each in 0.1% TBST, anti-mouse secondary antibody (Abcam, UK—Cat. No. ab6728; 1:5000 dilution in 0.1% TBST) was added to the membrane and incubated for 1 hour at room temperature, with rocking. The membrane was then washed three times with 0.1% TBST, each for 5 minutes, at RT on a rocker-shaker. The blot was developed using an ECL reagent (Millipore, USA), according to manufacturer’s instructions. Uncropped blots are provided in the Supplementary Information.
4
Immunohistochemical Analysis of ER, GFP, and Kisspeptin
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Immunohistochemistry was performed on cryosections (20 µm) collected from brains fixed in 4% paraformaldehyde using standard procedures. ERα staining used rabbit polyclonal anti-ERα antibody (EMD Millipore, Billerica, MA cat # C1355) or mouse monoclonal ERα antibody (Abcam, Cambridge cat # 93021) at a dilution of 1:1000 or 1:100, respectively. GFP staining used anti-GFP antibody (Novus Biologicals, Littleton, CO cat # NB100-1614) at 1:2500. Kisspeptin staining used rabbit polyclonal anti-KISS1 at a dilution of 1:200 (Abcam, Cambridge cat # ab19028). Confocal images were taken using a Nikon Ti inverted fluorescence microscope with CSU-22 spinning disk confocal.
5
Immunoprecipitation and Immunoblotting Protocol
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Rabbit anti-hnRNPUL1 antibody was purchased from Proteintech (Chicago, IL). Mouse anti-FLAG (M2) antibody, anti-β-tubulin antibody, and protein-A-Sepharose beads were purchased from Sigma (St. Louis, MO). Anti-GFP antibody was purchased from Novus Biologicals (Littleton, CO). Rabbit anti-PRMT1 antibody and ASYM25b were described previously21 (link)36 (link). Immunoprecipitations and immunoblotting were performed as previously described37 (link). Briefly, cells were lysed in 50 mM HEPES pH 7.4, 150 mM NaCl, and 1% Triton X-100 on ice for 15 min. After removal of the Triton insoluble matter by centrifugation, the supernatant was incubated with the indicated antibodies on ice for 2 h. The bound proteins were immunopurified using protein A Sepharose beads tumbled at 4 °C for 1 h and separated by SDS-PAGE, transferred to nitrocellulose membranes and immunoblotted with the indicated antibodies, as previously described37 (link).
6
GFP Protein Detection by Western Blot
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Ten micrograms of protein for each sample was resolved on a 15% SDS-PAGE, transferred onto a PVDF membrane (Millipore, Billerica, MA) and blocked with 5% w/v skimmed milk protein prepared in 0.1% TBST (room temperature, 1 hr). Subsequently, the membrane was incubated overnight with anti-GFP antibody (Novus, USA -Cat. No. NB100-2220; 1:1000 dilution in 0.1% TBST) at 4 °C on a rocker-shaker. Following three washes of 5 minutes each in 0.1% TBST, anti-mouse secondary antibody (Abcam, UK -Cat. No. ab6728; 1:5000 dilution in 0.1% TBST) was added to the membrane and incubated for 1 hour at room temperature, with rocking. The membrane was then washed three times with 0.1% TBST, each for 5 minutes, at RT on a rocker-shaker. The blot was developed using an ECL reagent (Millipore, USA), according to manufacturer's instructions.
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