AAP was obtained from Psaitong (Beijing, China). AA was obtained from Sinopharm (Beijing, China). Colormixed protein marker and nickel-nitrilotriacetic acid (Ni-NTA) Sepharose were obtained from Solarbio (Beijing, China). OTA, ochratoxin B (OTB), ochratoxin C (OTC), zearalenone (ZEN), aflatoxin B1 (AFB1), fumonisin B1 (FB1), and deoxynivalenol (DON) standards were procured from Pribolab (Qingdao, China). 3,3′,5,5′-Tetramethyl benzidine (TMB), p-nitrophenylphosphate (pNPP), ovalbumin (OVA), bovine serum albumin (BSA), and 96-well microplates were obtained from Sangon Biotech (Shanghai, China). Tetramethylammonium hydroxide (TMA·OH) and manganese chloride tetrahydrate (MnCl2·4H2O) were purchased from Aladdin (Shanghai, China). Polyvinylidene fluoride membrane was obtained from Millipore (Bearrica, MA). OTA–BSA conjugates were prepared previously in the laboratory. The recombinant pET25b-Nb-G53Q&S102D vector containing the mutated Nb gene for OTA and the recombinant pecan45-Nb28-AP vector were prepared in previous works.24,33 (link)
Colormixed protein marker
Manufactured by Solarbio
Sourced in China
The ColorMixed Protein Marker is a pre-stained protein molecular weight standard designed for monitoring protein separation during SDS-PAGE. It contains a mixture of colored proteins with defined molecular weights, allowing for easy visualization and identification of protein bands.
Automatically generated - may contain errors
Lab products found in correlation
Ecl western blotting substrate, by Solarbio (3 mentions)
Sds page gel preparation kit, by Beyotime (2 mentions)
Paraformaldehyde (pfa), by Solarbio (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (2 mentions)
Tbst buffer, by Solarbio (2 mentions)
Methyl thiazolyl tetrazolium (mtt), by Solarbio (2 mentions)
Ripa lysis buffer, by Solarbio (2 mentions)
Lysotracker, by Beyotime (2 mentions)
Sds page sample loading buffer, by Solarbio (2 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Nickel nitrilotriacetic acid ni nta sepharose, by Solarbio (1 mentions)
Deoxynivalenol, by Pribolab (1 mentions)
96 well microplates, by Sangon (1 mentions)
Quantity one analysis software, by Bio-Rad (1 mentions)
R0010, by Solarbio (1 mentions)
P1200, by Solarbio (1 mentions)
Ya1701, by Solarbio (1 mentions)
P1045, by Beyotime (1 mentions)
Matrigel, by Corning (1 mentions)
2 7 dichlorodihydrofluorescein diacetate dcfh da, by Merck Group (1 mentions)
6 protocols using colormixed protein marker
1
Nanobody-Based Ochratoxin A Detection
2
Western Blot Protein Analysis Protocol
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The total protein was lysed from cells in radio immuno precipitation assay buffer (R0010, Solarbio) with protease and phosphatase inhibitor (P1045, Beyotime, 1:50) for 30 min at 4°C, and then quantified with a BCA kit (PC0020, Solarbio). ColorMixed Protein Marker (PR1920, Solarbio) served as a protein size marker. Then, protein (20 μg) was isolated by 6–10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis gels (P1200, Solarbio) and transferred onto immobilon-P polyvinylidene fluoride membranes (YA1701, Solarbio). Following incubation with 5% bovine serum albumin (SW3015, Solarbio) for 1 h at room temperature, the membranes were treated with primary antibodies (shown in Table 2 ) at 4℃ overnight, and then corresponding secondary antibody (shown in Table 2 ) for 2 h at room temperature. Visualization was achieved with ECL Western Blotting Substrate (PE0010, Solarbio). The intensity of protein band was determined by Quantity One Analysis Software (version 4.62; Bio-Rad Laboratories, Inc.).
3
Nanoparticle-mediated Delivery of Angiogenic Factors
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pVEGF and pNGF were purchased from Aibimeng Biotechnology Co., Ltd (Jiangsu, China). Protamine sulfate and coumarin 6 were obtained from Aladdin (Shanghai, China). Paraformaldehyde, DAPI, MTT, RIPA Lysis Buffer, 5 × SDS-PAGE Sample Loading Buffer, 20 × TBST buffer, ECL Western Blotting Substrate and Color Mixed Protein Marker were purchased from Solarbio (Beijing, China). SDS-PAGE gel preparation kit and Lyso-Tracker were bought from Beyotime (Jiangsu, China). Hydrogen peroxide (H2O2, 30 wt %) was purchased from Tianjin Guangfu Fine Chemical Research Institute (Tianjin, China). 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) was purchased from Sigma-Aldrich (St.Louis, MO, USA). Matrigel was purchased from Corning (New York, USA).
4
Polysaccharide Extraction from Blackberry
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Chicken breast was purchased from Fujian Chia Tai Food Co. Ltd (Longyan city, China), and the powder of blackberry (Rubus spp.) crude polysaccharide was extracted in laboratory and stored at 4 °C until use. ColorMixed Protein Marker, Coomas bright blue quick dye solution, urea, Tris solution, and SDS solution were ordered from Solarbio Science & Technology Co. Ltd (Beijing, China). Other chemicals including EDTA and potassium bromide were purchased from Macklin biochemical Co. Ltd. (Shanghai, China).
5
Cobalt (II) Chloride Hexahydrate Protocol
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Cobalt (II) chloride hexahydrate was purchased from Sigma (St. Louis, MO, USA); Anti–HIF–1α, Anti-beta Actin, IgG H&L (DyLight 488) and IgG H&L (HRP) were obtained from Abcam (Cambridge, UK); Paraformaldehyde, DAPI, MTT, RIPA Lysis Buffer, 5 × SDS-PAGE Sample Loading Buffer, 20 × TBST buffer, ECL Western Blotting Substrate and Color Mixed Protein Marker were from Solarbio (Beijing, China); SDS-PAGE gel preparation kit and Lyso-Tracker were bought from Beyotime (Jiangsu, China).
6
Western Blot Analysis of Protein Targets
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Total protein was extracted by RIPA Buffer (89900, Thermo Fisher, U.S.A.) and quantified with BCA kits (A53227, Thermo Fisher, U.S.A.). The harvested protein samples with the protein-loading buffer (P0015, Beyotime) were boiled for 5 min, and loaded into 10% SDS/PAGE gel for electrophoresis separation with ColorMixed Protein Marker (11–180 kDa, PR1910, Solarbio). Then, the protein was transferred on to PVDF membrane (YA1701, Solarbio) and blocked with 5% bovine serum albumin (BSA) (SW3015, Solarbio) at 37°C for 2 h. The membrane was incubated firstly with the primary antibodies (DUSP1 (#48625, Rabbit, 1:1000, CST, U.S.A.), NOR-1 (ab155535, Rabbit, 1:3000, Abcam, Cambridge, MA, U.S.A.), PCNA (ab29, Mouse, 1:1000, Abcam), GAPDH (Mouse, ab8245, 1:10000, Abcam)) at 4°C overnight, and subsequently with the secondary antibody (Goat Anti-Rabbit IgG H&L (HRP) (ab205718, 1:20000, Abcam) or Goat Anti-Mouse IgG H&L (HRP) (ab205719, 1:20000, Abcam)) for 1 h. Then, the membrane was developed by the BeyoECL Moon (P0018FS, Beyotime) in the dark. The protein signal was detected using Bio-Rad gel imaging system (ChemiDocXRS + Imaging System), with GAPDH serving as the internal reference. The original Western blot images were shown in Supplementary Figures S1 and S2.
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