For Nischarin/PAK binding, cortical neurons were lysed in RIPA lysis buffer with protease and phosphatase inhibitors. Cell lysates (500 μl, concentration 3 μg/μl) were incubated with anti-Nischarin antibody, anti-PAK1 or anti-PAK2 antibody at 4°C for 6 h. For controls, cell lysates were incubated with the same amount of immunoglobulin G (IgG, Sigma). After incubation with Protein G-Sepharose beads for 2 hours, the immunoprecipitates were washed three times with ice-cold lysis buffers, and the binding proteins were eluted with sample buffer. The general procedures for western blot were the same as described above.
Immunoglobulin g igg
Manufactured by Merck Group
Sourced in United States
Immunoglobulin G (IgG) is a type of antibody found in the blood and body fluids. It is the most abundant antibody class in the human body, comprising approximately 75% of all antibodies. IgG plays a critical role in the adaptive immune response, binding to pathogens and marking them for destruction by other components of the immune system.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Magna rip rna binding protein immunoprecipitation kit, by Merck Group (2 mentions)
Exonuclease 1, by New England Biolabs (1 mentions)
Milli q water purification system, by Merck Group (1 mentions)
Sertraline, by Merck Group (1 mentions)
Fluoxetine, by Merck Group (1 mentions)
Hemoglobin, by Merck Group (1 mentions)
Fibronectin, by Merck Group (1 mentions)
Glutamine, by Merck Group (1 mentions)
Human serum, by Merck Group (1 mentions)
Aunps, by Merck Group (1 mentions)
Sodium dodecyl sulphate (sds), by Merck Group (1 mentions)
Sodium dihydrogen phosphate, by Merck Group (1 mentions)
Cytochrome c cyt c, by Merck Group (1 mentions)
Hepes, by Merck Group (1 mentions)
Tween 20, by Merck Group (1 mentions)
Glycerol, by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Urea, by Fagron (1 mentions)
14 protocols using immunoglobulin g igg
1
Nischarin/PAK Binding in Cortical Neurons
2
Sensitive DNA Detection Using Fluorescence
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The DNA sequences are as follows. P1: 5′-CGACATCTAACCTAGCTGACT-3′ P2: 5′-AGTCAGCTAGGTTAGATGTCG-3′. The two oligonucleotides, P1 and P2, were purchased and purified from Sangon Biological Engineering Technology & Services Co., Ltd. (Shanghai, China). Streptavidin (SA), Carcinoembryonic antigen (CEA), alpha fetoprotein (AFP) and Immunoglobulin G (IgG) were all obtained from Sigma-Aldrich (St. Louis, MO, USA). The ultrapure water (18.2 MΩ.cm) used in the experiments was from a Milli-Q water purification system (Millipore Corp, Bedford, MA, USA). The reaction buffer in the experiment system was Tris-HCl buffer (20 mM Tris-HCl, 150 mM NaCl, 10 mM MgCl2, pH7.5). Tris [Tris- (hydroxy- methyl) aminomethane], hydrochloric acid (HCl), sodium chloride (NaCl) and agnesium chloride (MgCl2) were purchased from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Exonuclease I were obtained from New England Biolabs (Beverly, MA, USA). SYBR green I (20×) was purchased from ZeesanBiotech co., Ltd. (Xiamen, China). Poly[(9,9-bis(6′-N,N,N-triethy-lammonium)hexyl) fluorenylene phenylene dibromide] (PFP) was purchased from Yuanye Co., Ltd. (Shanghai, China).
3
Diffusion and Permeability of Small Molecules in Placental Transport
4
Degradation of Human Proteins by rAcCP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess the degradation of human proteins by rAcCP, various proteins (2 mg/ml) were separately incubated with rAcCP (100 nM) in 50 mM sodium acetate (pH 4.0) or Tris-HCl (pH 7.0) in the presence of 1 mM GSH for 3 hr at 37°C. The proteins used for hydrolysis assay were hemoglobin (from human blood), albumin (from human serum), immunoglobulin G (IgG; from human serum), IgA (from human serum) and fibronectin (from human plasma), all purchased from Sigma. After the reactions were terminated by adding reducing sample buffer, protein degradation was analyzed by SDS-PAGE.
5
LN Podocyte Autophagy Inhibition Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human immortalized podocytes were obtained from American Type Culture Collection and cultured in RPMI-1640 medium (Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 10% fetal bovine serum (FBS; Thermo Fisher Scientific) and 2 mM glutamine (Sigma, St. Louis, MA, USA) at 37°C. Podocytes were treated with immunoglobulin G (IgG; Sigma) for 24 h to establish an in vitro model of LN. To confirm the correlation between Tim-1 and autophagy in LN, podocytes were treated with an autophagy inhibitor (3-methyladenine and bafilomycin A1, Sigma).
6
Aptamer-based Detection of Cardiac Troponin I
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cTnIs were added to a hybridization buffer solution (300 mM NaCl, 20 mM Tris-HCl, 0.5 mM beta-mercaptoethanol, 20% (w/v) glycerol, 0.05% Tween 20, pH 8.0) or a hybridization buffer solution with 20% human serum (Sigma-Aldrich). Au NPs (~10 nm) were purchased from Sigma-Aldrich and used after centrifugation (10,000× g, 30 min). Au NPs were dispersed in a final 500 µL of 1 µM reporter aptamer solution (0.05 M PBS, 5 mM NaCl, 2.5 mM KCl, 0.5 mM MgCl2, pH 7.4) at room temperature for 3 h. To remove excess reporter aptamers, the Au NP solution was centrifuged (10,000× g, 15 min), the supernatant was removed, and resuspended in PBS (500 µL). For the detection of cTnIs, the prepared cTnI solution (1 mL) was added to the aptamer-immobilized Au nanoplates at 35 °C. After 6 h, the Au nanoplates were washed with the hybridization buffer and distilled deionized water. Next, the cTnI-captured Au nanoplates were incubated in the Au NP reporter solution at 35 °C for 30 min. Finally, the cTnI-detected Au NP-on-nanoplate structures were rinsed with deionized water and dried by N2 gas. The same experimental procedure was accomplished using troponin C (TnC, Abcam), troponin T (TnT, Sigma-Aldrich), immunoglobulin G (IgG, Sigma-Aldrich), and avidin (Sigma-Aldrich) instead of cTnI for the selectivity test.
7
Synthesis of Lactose and Sulfated Lactose
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The two disaccharides (Scheme 1) lactose (BB1) and sulfated lactose (BB2) were prepared from a βlactoside, which was first converted to lactosylated disulfide BB1. Then, sulfatation was performed to obtain the fully sulfated disulfide BB2. The detailed synthesis of the disaccharides was reported previously. [35] Sodium chloride, sodium dihydrogen phosphate, HEPES, sodium dodecyl sulphate (SDS), Tween 20 and glycerol were purchased from Sigma-Aldrich. All of them were used as received. Protein lectin from Erythrina cristagalli (ECL), bovine serum albumin (BSA), Immunoglobulin G (IgG) from rabbit serum, and Cytochrome C (CytC) from horse heart were also purchased from Sigma-Aldrich and used as received. In this study, ultrapure water (18.2 MΩ.cm) used for preparation of buffer solutions was produced by a Purelab classic system (Elga).
8
Electrochemical Biosensor Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All chemicals were of analytical grade and ultrapure Milli-Q water laboratory grade (conductivity b0.1 μS/cm) was used. The chemical reagents used included potassium hexacyanoferrate (III) (K 3 [Fe(CN) 6 ]) and potassium hexacyanoferrate (II) trihydrate (K 4 [Fe(CN) 6 ].3H 2 O), obtained from Riedel-de Häen; Phosphate buffered saline (PBS) tablets purchased from Sigma-Aldrich; 3,4-Ethylenedioxythiophene, 97% (EDOT), from Alfa Aesar; 4-Aminothiophenol, 96% (4-AMP), from Acros Organics; EriochromeBlack T (EBT), from Biochem; Interleukin-1β (IL-1β) (AA 117-269) with purity N95% in PBS pH 7.4 (concentration 1 mg/mL) from Antibodies-online, Germany. Myoglobin (Myo) and Immunoglobulin G (IgG) were purchased from Sigma-Aldrich; Glucose (Glu) from Fisher BioReagents and Urea from Fagron; sodium chloride (NaCl) from Panreac; potassium chloride (KCl) and calcium chloride (CaCl 2 ) from Merck; magnesium chloride hexa-hydrate (MgCl 2 .6H 2 O) from Riedel-de Häen was also used.
9
ChIP Assay for Histone Modifications
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The ChIP assay was performed using the EZ-Magna ChIP A/G Kit (Millipore) according to the manufacturer’s instructions. Cells were fixed and centrifuged, and the pellets were resuspended in ChIP lysis buffer. After sonication, the sheared DNA complex of nuclear lysates was collected, and antibodies were added, including H3K27me3, SUZ12, EZH2 (Active Motif), RNA polymerase II, and immunoglobulin G (IgG) (Millipore). The mixture was incubated overnight, and magnetic beads were added to pull down the DNA-protein-antibody complexes. After elution, the samples were ready for real-time PCR analysis. The primers for ChIP-qPCR are listed in Table S1 .
10
Investigating miR-1253 and hsa_circ_0000190 Interaction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To confirm the ability of miR-1253 and hsa_circ_0000190 to interact directly with one another, the EZ-Magna RIP™ RNA-Binding Protein Immunoprecipitation Kit (Millipore) kit was used based on provided instruction. Lysis buffer supplemented with RNase inhibitor (Millipore) was used for the initial preparation of cell lysates, which were subsequently incubated with Argonaute2 (Ago2; Millipore) or Immunoglobulin G (IgG; Millipore) antibody-coated magnetic beads. A qPCR approach was subsequently used to quantify RNA enrichment.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!