Tissues were homogenized in lysis buffer (50 mM Tris-HCl pH 7.4, 250 mM NaCl, 0.5% (vol/vol) Igepal with complete protease and phosphatase inhibitors [protease inhibitors: 0.1 μm aprotonin (USB), 0.02 mM leupeptin (USB), 0.01 mM pepstatin (USB), 0.5 mM PMSF (Sigma); phosphatase inhibitors: 2 mM imidazole (Sigma), 1.15 mM sodium molybdate (Sigma), 1 mM sodium orthovanadate (Sigma), 5 nM microcystin (Calbiochem)]. Western blotting was performed with the following antibodies from: Cell Signaling Technology: BAP1 (#13271); Bethyl Laboratories: PBRM1 (A301-591A), Hif-1α (A300-286A); Novus Biologicals: CAIX (AF2344), Hif-2α (AF2997); Santa Cruz Biotechnology: VHL (sc-1534); Millipore: H2A (07-146), Ubiquityl-Histone H2A (05-678); Sigma: Tubulin (T5168); Thermo Fisher Scientific: HRP-conjugated goat anti-mouse IgG (#31430) and HRP-conjugated goat anti-rabbit IgG (#31460); Novus Biologicals: Donkey anti-Goat IgG Secondary Antibody (HAF109).
Hrp conjugated goat anti mouse igg
Manufactured by Novus Biologicals
Sourced in Japan
HRP-conjugated goat anti-mouse IgG is a secondary antibody that binds to mouse immunoglobulin G (IgG) and is conjugated to horseradish peroxidase (HRP). This facilitates the detection of mouse IgG in various immunoassays and immunochemical techniques.
Automatically generated - may contain errors
Lab products found in correlation
Sodium molybdate, by Merck Group (1 mentions)
Imidazole, by Merck Group (1 mentions)
Hrp conjugated goat anti rabbit igg, by Novus Biologicals (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Merck Group (1 mentions)
Sodium orthovanadate, by Merck Group (1 mentions)
Moab 2, by Novus Biologicals (1 mentions)
Hrp conjugated goat anti mouse igg, by Boster Bio (1 mentions)
4 protocols using hrp conjugated goat anti mouse igg
1
Protein Extraction and Western Blotting
2
Binding Affinity Measurement of Monoclonal Antibodies
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Purified proteins were used to coat 96-well plates (30 ng/well) overnight at 4°C. Culture supernatants from individual hybridomas were added to each well (100 µl/well) followed by horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Novus). The values of the optical density (OD) of substrate reactions were read at 450 nm on a plate reader (Bio-Rad).
The binding ability of mAb was measured by ELISA as described previously [31 (link),32 (link)]. MAbs were serially two-fold diluted at an initial concentration of 10 µg/ml and detected by ELISA as described above.
The binding ability of mAb was measured by ELISA as described previously [31 (link),32 (link)]. MAbs were serially two-fold diluted at an initial concentration of 10 µg/ml and detected by ELISA as described above.
3
Binding Affinity Measurement of Monoclonal Antibodies
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Purified proteins were used to coat 96-well plates (30 ng/well) overnight at 4°C. Culture supernatants from individual hybridomas were added to each well (100 µl/well) followed by horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG (Novus). The values of the optical density (OD) of substrate reactions were read at 450 nm on a plate reader (Bio-Rad).
The binding ability of mAb was measured by ELISA as described previously [31 (link),32 (link)]. MAbs were serially two-fold diluted at an initial concentration of 10 µg/ml and detected by ELISA as described above.
The binding ability of mAb was measured by ELISA as described previously [31 (link),32 (link)]. MAbs were serially two-fold diluted at an initial concentration of 10 µg/ml and detected by ELISA as described above.
4
Immunostaining Protocol for IgG and Aβ
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Immunostaining for IgG was carried out as previously described [26 (link), 27 (link)]. Briefly, the 30 μm-thick sections were washed in 0.01 M PBS three times in order to wash out cryoprotectant and then incubated in 3% hydrogen peroxide (methanol: 30% H2O2 = 1:1 in 0.01 M PBS) for 20 min, permeabilized with PBS containing 0.25%Triton X-100 (PBST, 3 × 10 min). The sections were blocked in 5% normal goat serum at RT for 1 h, followed incubation with HRP-conjugated goat anti-mouse IgG (Boster Biological Technology, Wuhan, China) overnight at 4 °C. Free-floating sections were subsequently washed in PBS three times. Sections were incubated in 0.05% diaminobenzidine (DAB) to visualize reaction products and then washed with PBS (3 × 5 min), mounted to a glass slide. Then sample was dried in air overnight, dehydrated using graded ethanol, cleared in xylene before cover-slipped with Neutral balsam. Images were captured under a light microscope (Olympus Corporation, Tokyo, Japan).
Immunostaining for Aβ was followed with the above procedures, incubated with primary antibody MOAB2 (Novus) followed by HRP-conjugated goat anti-mouse IgG secondary antibody.
Immunostaining for Aβ was followed with the above procedures, incubated with primary antibody MOAB2 (Novus) followed by HRP-conjugated goat anti-mouse IgG secondary antibody.
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