In the experimental procedures described, the following antibodies were used: rabbit monoclonal anti-LRP1 (Ab92544; Abcam), mouse monoclonal anti-ABCA1 (Ab18180; Abcam), rabbit monoclonal anti-PSAP (ab166910; Abcam), mouse monoclonal anti-CTSD (NBP1-04278 Novus Biologicals), goat anti–SR-B1 (NB400-131; Novus Biologicals), rabbit anti-ABCG1 (ab36969; Abcam), rat anti–Wnt-5a (MAB645; R&D Systems), mouse monoclonal anti–β-actin (A5441, Sigma-Aldrich, 1:5,000), anti–α-tubulin (Ab4047, Abcam), anti-GAPDH (Ab8245), rabbit anti-GST (sc-459), goat anti-rabbit IgG-horseradish peroxidase conjugate (170–6515: Bio-Rad Laboratories,), goat anti-mouse IgG-horseradish peroxidase conjugate (170-6516; Bio-Rad Laboratories), and goat anti-Rat IgG-horseradish peroxidase conjugated (HAF005; R&D Systems). For immunofluorescence, Alexa Fluor–conjugated secondary antibodies and Alexa Fluor 488–conjugated and 568–conjugated antibodies were purchased from Jackson ImmunoResearch.
Goat anti mouse igg horseradish peroxidase conjugate
Manufactured by Bio-Rad
Sourced in United States
Goat anti-mouse IgG-horseradish peroxidase conjugate is a laboratory reagent that consists of goat-derived antibodies specific to mouse immunoglobulin G (IgG), coupled to the enzyme horseradish peroxidase. This conjugate is commonly used as a detection tool in various immunoassay techniques.
Automatically generated - may contain errors
Lab products found in correlation
Nitrocellulose membrane, by GE Healthcare (2 mentions)
A5441, by Merck Group (1 mentions)
Ab166910, by Abcam (1 mentions)
Nbp1 04278, by Novus Biologicals (1 mentions)
Ab8245, by Bio-Rad (1 mentions)
Goat anti rat igg horseradish peroxidase conjugated haf005, by R&D Systems (1 mentions)
Mab645, by R&D Systems (1 mentions)
Nb400 131, by Novus Biologicals (1 mentions)
Alexa fluor conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Ab92544, by Abcam (1 mentions)
Ab4047, by Abcam (1 mentions)
Ab36969, by Abcam (1 mentions)
Ab18180, by Abcam (1 mentions)
Goat anti rabbit igg horseradish peroxidase conjugate, by Bio-Rad (1 mentions)
Goat anti rabbit igg, by EASYBIO (1 mentions)
Opteia, by BD (1 mentions)
Ecl prime, by GE Healthcare (1 mentions)
Anti dc stamp mab clone 1a2, by BD (1 mentions)
Xt mes electrophoresis buffer, by Bio-Rad (1 mentions)
Anti rabbit igg horseradish peroxidase conjugate, by Bio-Rad (1 mentions)
6 protocols using goat anti mouse igg horseradish peroxidase conjugate
1
Immunoblotting and Immunofluorescence Antibodies
2
Western Blot Analysis of Recombinant Proteins
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Total proteins were isolated from N. benthamiana leaf tissues in extraction buffer (100 mM Tris-HCl, pH 6.8, 20% glycerol, 4% SDS, 0.2% bromphenol blue, 5% β-mercaptoethanol). Total proteins were separated in a 4–15% SDS-PAGE gradient and transferred to nitrocellulose membrane (GE Healthcare Life Sciences). Membranes were blocked with 5% (m/v) skimmed milk powder at room temperature for 1 hr and then incubated with primary antibodies at 37°C for 1 hr. After washing three times, membranes were incubated with secondary antibodies at 37°C for 1 hr. Antibodies against RFP (1:3000), P (1:3000), GST (1:5000), GFP (1:5000; MBL, 598), Flag (1:5, 000; Sigma, F1804) were used for protein detection. Goat anti-rabbit IgG (EASYBIO, BE0101) and goat anti-mouse IgG horseradish peroxidase conjugate (Bio-Rad, 170–6516) were used as secondary antibodies. After addition of NcmECL Ultra stabilized peroxide reagent (NCM Biotech, P10300B), chemiluminescence of membranes was detected with a Biomolecular Imager (Azure biosystems, Inc).
3
ELISA-based VHH Fragment Detection
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Maxisorp® 96‐well plates were coated with purified m. pasudotox (with either the amidated form or with a ≈80% deamidated form at 5 sμg/mL in PBS (100 µL/well) and incubated for 1 h at RT followed by a 1 h blocking step with 2% P‐PBS (200 sμL/well). Crude E. coli supernatants with soluble VHH fragments were diluted five times in 1% P‐PBST and incubated on the coated plates (100 sμL/well) for 1 h at RT followed by a washing step using PBST. All following incubations were carried out at 100 sμL/well for 1 h at RT. The bound VHH fragments were detected using a mouse anti c‐Myc monoclonal antibody (Thermo Fisher Scientific) at 0.5 sμg/mL in 1% P‐PBST followed by incubation with a goat anti‐mouse‐IgG horseradish peroxidase conjugate (Bio‐Rad Laboratories) diluted 1:2000 in 1% P‐PBST. After washing of the plates , a color reaction was allowed using a 3,3',5,5'‐Tetramethylbenzidine/Ureaperoxide solution (OptEIA™; Becton Dickinson, Franklin Lakes, NJ, USA) at 100 sμL/well, which was stopped after 5–10 min by addition of 50 sμL/well 1 M H2SO4. Plates were measured in a plate reader at 450 nm. The same approach was taken for the anti‐prothrombin VHH fragments, except that the plates were coated with purified, fully gamma‐carboxylated human plasma‐derived prothrombin (Haematologic Technologies Inc.).
4
Analyzing DC-STAMP Protein Expression
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Reagent-treated cells were cultured in a 6-well plate (4 × 104 cells/well) for 72 h. The cells were washed twice with PBS before collection followed by dissolved in certain quantity of lysis buffer. The cell lysates were loaded onto a 10% gel for SDS-PAGE and blotted onto a PVDF membrane using a wet-transfer method. The membrane was blocked with 1% bovine serum albumin in TBS with 0.05% tween 20 (TBST) for 1 h, and then incubated at 4 °C for overnight with an anti-DC-STAMP mAb (clone 1A2, 1:500, Becton Dickinson) as the primary antibody. Goat anti-mouse IgG horse-radish peroxidase conjugate (1:3000, Biorad, Hercules, CA, USA) was used as the secondary antibody at room temperature for 1 h. The membrane was incubated with ECL prime (GE Healthcare, Chicago, IL, USA) at room temperature for 5 min in accordance with the manufacturer's protocol, and LAS imaging system (Wako, Osaka, Japan) was used for imaging.
5
Comprehensive Cell Culture Reagent Catalog
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Cell culture flasks and plates were purchased from Sarstedt (Nümbrecht, Germany) or TPP (Transandigrn, Switzerland). Advanced MEM, DMEM, OptiMEM, GlutaMAX, MEM vitamin solution, fetal bovine serum (FBS), Fungizone (250 µg/mL of amphotericin B), gentamicin (10 mg/mL), trypsin-EDTA, Pierce Enhanced Chemiluminescence (ECL) Western Blotting substrate, High Capacity cDNA Reverse Transcription Kit, TaqMan Universal Master Mix with UNG and TaqMan gene expression assays were purchased from Thermo Fisher Scientific (Waltham, MA, U.S.). 4-12% Criterion Bis-Tris polyacrylamide gels, XT MES electrophoresis buffer, goat anti-mouse IgG-horseradish peroxidase conjugate, anti-rabbit IgG-horseradish peroxidase conjugate, PCR plates and PCR plate sealing films were purchased from Bio-Rad (Hercules, CA, U.S.). Amersham ECL Full-Range Rainbow Molecular Weight Marker was purchased from (Cytiva, U.S.). Polyvinylidene fluoride (PVDF) membrane, forskolin, dibutyryl-cAMP (db-cAMP), bovine serum albumin (BSA), and all other reagents were purchased from Merck/Sigma-Aldrich (Darmstadt, Germany). Insulin (Actrapid) was obtained from NovoNordisk (Denmark). E.Z.N.A HP Total RNA Kit was purchased from Omega Bio-Tek (Norcross, GA, U.S.). Dexamethasone was purchased from KRKA (Slovenia).
6
Western Blot Analysis of Recombinant Proteins
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Total proteins were isolated from N. benthamiana leaf tissues in extraction buffer (100 mM Tris-HCl, pH 6.8, 20% glycerol, 4% SDS, 0.2% bromphenol blue, 5% βmercaptoethanol). Total proteins were separated in a 4-15% SDS-PAGE gradient and transferred to nitrocellulose membrane (GE Healthcare Life Science). Membranes were blocked with 5% (m/v) skimmed milk powder at room temperature for 1 h and then incubated with primary antibodies at 37°C for 1 h. After washing three times, membranes were incubated with secondary antibodies at 37°C for 1 h. Antibodies against RFP (1:3000), P (1:3000), GST (1:5000), GFP (1:5000; MBL, 598), Flag (1:5, 000; Sigma, F1804) were used for protein detection. Goat anti-rabbit IgG (EASYBIO, BE0101) and goat anti-mouse IgG horseradish peroxidase conjugate (Bio-Rad, 170-6516) were used as secondary antibodies. After addition of NcmECL Ultra stabilized peroxide reagent (NCM Biotech, P10300B), chemiluminescence of membranes was detected with a Biomolecular Imager (Azure biosystems, Inc.).
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