An aliquot of the extract (500 μL) from Sf9 cells co-expressing the FLAG- and c-Myc-tagged receptors was supplemented with a 50% slurry (20 µL) of agarose-conjugated anti-FLAG anti-body (Santa Cruz Biotechnology, Inc.). The mixture was shaken overnight at 4°C, and immunoadsorbed receptor was collected by centrifugation. The precipitated beads then were washed 4 times with 3 mL of buffer C (20 mM HEPES, 1 mM EDTA, 0.1 mM PMSF, adjusted to pH 7.40 with NaOH) supplemented with digitonin (1%) and cholate (0.001%), and the entire precipitate was applied to the polyacrylamide gel. Following electrophoresis and transfer, the nitrocellulose membrane (Bio-Rad, 0.45 µm) was blotted with anti-c-Myc antibody (Santa Cruz Biotechnology, Inc.) as described previously (Ma et al., 2007 (link)).
Anti c myc antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Anti-c-Myc antibody is a laboratory research tool used to detect and study the c-Myc protein, which is a transcription factor involved in cell growth and proliferation. This antibody can be used in various immunoassays, such as Western blotting, immunoprecipitation, and immunofluorescence, to identify and analyze the c-Myc protein in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Nitrocellulose membrane, by Bio-Rad (3 mentions)
Anti β actin antibody, by Merck Group (3 mentions)
Anti β catenin antibody, by Santa Cruz Biotechnology (3 mentions)
Lysis buffer, by Cell Signaling Technology (3 mentions)
O phenylenediamine dihydrochloride reagent, by Merck Group (2 mentions)
Horseradish peroxidase hrp conjugated goat anti rabbit igg, by Cell Signaling Technology (2 mentions)
Image station 2000 mm, by Kodak (2 mentions)
Molecular imaging software version 4, by Kodak (2 mentions)
Anti bcl 2 antibody, by Santa Cruz Biotechnology (1 mentions)
Anti bax antibody, by Santa Cruz Biotechnology (1 mentions)
Anti caspase 3 antibody, by Santa Cruz Biotechnology (1 mentions)
Complete mini, by Roche (1 mentions)
Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions)
Immobilon p membrane, by Merck Group (1 mentions)
Supersep gel, by Fujifilm (1 mentions)
Anti rabbit igg antibody, by Promega (1 mentions)
Anti cyp b, by Thermo Fisher Scientific (1 mentions)
Horseradish peroxidase conjugated anti mouse igg antibody, by Promega (1 mentions)
Anti pin1 antibody, by Cell Signaling Technology (1 mentions)
Anti cyclin d1, by Santa Cruz Biotechnology (1 mentions)
36 protocols using anti c myc antibody
1
Immunoprecipitation and Western Blot Analysis
2
Immunohistochemical Analysis of c-Myc in HCC
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In this study, IHC staining of the HCC samples was performed according to the protocol described previously.26 (link) Xylene was used to dewax the paraffin-embedded tissue sections, and the deparaffinized sections were rehydrated using ethanol. Then, epitope retrieval was induced by in citrate buffer (pH 6.0). Anti-c-Myc antibody was used to identify C-Myc expression (Santa Cruz, CA, USA).
3
Molecular Mechanisms of Cardiac Remodeling
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The experimental protocol was approved by the Animal Care and Use Committee of Xuzhou Medical College. Rats were housed in a climate-controlled room. Sterile water and standard chow diet were available ad libitum. Six-week-old male Sprague-Dawley (SD) rats were provided by the Animal Department, Xuzhou Medical College. All of the antibodies, including anti-forkhead box H1 (Foxh1) antibody, anti-c-Myc antibody, anti-p-Smad3 antibody, anti-matrix metalloproteinase 2 (MMP2) antibody, anti-tissue inhibitor of matrix metalloproteinases type 2 (TIMP2) antibody, anti-Bax antibody, anti-Bcl-2 antibody, anti- Caspase-3 antibody and anti-APJ antibody, were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). All other reagent-grade chemicals were purchased from the Sigma- Aldrich Chemical Co. (St. Louis, MO, USA).
4
Western Blot Analysis of Coronavirus Infection
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The cell membranes were disrupted with cell lysis buffer [10 mM Tris–HCl, pH 7.8, 1 mM ethylenediamine tetraacetic acid (EDTA), 1% NP-40, and 0.15 M NaCl], including Complete Mini (Roche Diagnostics, Tokyo, Japan) at 20 h after infection. The cell lysates were resolved by electrophoresis on 12.5% SuperSep gels (WAKO, Tokyo, Japan) and Western blotted on to Immobilon-P membranes (Millipore, Tokyo, Japan). Non-specific protein binding was blocked with 5% non-fat dry milk, and then the membranes were incubated with the primary antibodies [anti-FCoV nucleocapsid (N) antibody (FIPV3-70; MyBioSource, CA, USA), anti-c-Myc antibody (Santa Cruz Biotechnology, CA, USA), anti-Pin1 antibody (Cell Signaling Technology, Tokyo, Japan), anti-Cyp B (Thermo Fisher Scientific, Yokohama, Japan), and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH; Calbiochem, CA, USA)] for 1 h. Antigen signals were visualized by reacting proteins on the membranes with horseradish peroxidase-conjugated anti-mouse IgG antibody (Promega) and/or anti-rabbit IgG antibody (Promega) followed by an enhanced chemiluminescence substrate (SuperSignal West Femto Maximum Sensitivity Substrate; Thermo Fisher Scientific) according to the manufacturer's protocol.
5
Western Blot Analysis of Protein Targets
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Protein from tissue and cells was separated in a 12 % SDS-PAGE gel and transferred onto a nitrocellulose membrane (Bio-Rad, Hercules, USA). The membrane was blocked with 5 % non-fat milk and incubated with anti-c-Myc antibody, anti-Cyclin D1 (Santa Cruz, CA) or anti-β-actin antibody (Sigma, CA, USA). After being washed extensively, a goat anti-mouse secondary antibody (Pierce, IL, USA) was added to the system. The proteins were detected using ECL reagents (Pierce).
6
Immunofluorescence Assay for hLH-R
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IF was performed on Hec1A cells previously transfected with the vector containing the mogpLuc2AhLH-R sequence. The anti c-myc antibody (4 μg/ml, Santa Cruz Biotechnology) was used as primary antibody, and the anti-mouse Alexa488 (1 μg/ml, Invitrogen, Thermo Fisher) as secondary antibody. Images were acquired with Nikon D-Eclipse C1 (Nikon) confocal microscope.
7
Western Blot Analysis of Protein Targets
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Cells were lysed with RIPA Lysis Buffer (P0013B, Beyotime, China) or Nuclear and Cytoplasmic Protein Extraction Kit (P0027, Beyotime, China). Then protein concentrations were quantified with BCA Protein Assay Kit (P0011, Beyotime, China). Twenty microlitre of proteins were separated by SDS-PAGE and then transferred onto PVDF membranes. After blocking, the membranes were incubated with the one of following antibodies: anti-CARMA3 antibody (1:1000, bs-7081R, Bioss, China), anti-MMP2 antibody (bs-0412R, Bioss), anti-MMP9 antibody (1:1000, bs-4593R, Bioss, China), anti-β-catenin antibody (1:1000, #8480, Cell Signaling Technology, USA), anti-Survivin antibody (1:1000, #2808, Cell Signaling Technology, USA), anti-C-myc antibody (1:500, sc-40, Santa Cruz Biotechnology, USA), anti-beta-Actin (1:5000, bsm-33036 M, Bioss, China), or anti-Histone H3.1 (1:500, bs-17422R, Bioss, China) at 4 °C overnight. Then the membranes were incubated in HRP-labeled Goat Anti-Rabbit IgG (H + L) (1:5000, A0208, Beyotime, China) or HRP-labeled Goat Anti-Mouse IgG (H + L) (1:5000, A0216, Beyotime, China) at 37 °C for 45 mins. After incubating ECL reagent for 5 mins, the proteins were detected by ECL Select Western Blotting Detection System (Beyotime, China).
8
Chromatin Immunoprecipitation (ChIP) Assay Protocol
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The ChIP assay was performed using the EZ ChIP™ Chromatin Immunoprecipitation Kit from Millipore according to the manufacturer’s instructions. Briefly, after cells underwent gene transfection and treatment, cells were subjected to cross-linking of the chromatin, sonication and then immunoprecipitation of DNA-protein complex using an anti-c-Myc antibody (Santa Cruz Biotechnology) according to the kit’s protocol. After that, the immunoprecipitated DNA-protein complex was extracted for DNA and amplified using qPCR according to a standard qPCR protocol.26 (link)
9
Extraction and Quantification of mRNA
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Cell extracts were prepared as described above with lysis buffer containing 100 units/ml RNasin (Promega) and then diluted to a concentration of approximately 5 mg/ml. Input samples were collected at this point. Diluted extracts were incubated with anti-c-Myc antibody (Santa Cruz, SC-40) for 2 h at 4°C, followed by addition of protein A-Sepharose beads (GE healthcare, #17513801). After incubation for 2 h, beads were washed five times with lysis buffer. RNA was extracted from immunoprecipitates and 3% of input samples using TRIzol (Invitrogen) following the manufacturer’s instructions. cDNA was synthesized using oligo(dT) primers, and qRT-PCR was performed on Quantstudio 3. mRNA levels of immunoprecipitation samples were normalized to input samples.
10
Western Blot Analysis of Recombinant Proteins
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The purified recombinant proteins (4 µg/lane) were subjected to SDS-PAGE using 10% gels and then transferred to polyvinylidene difluoride membranes. Non-specific binding was blocked for 1 h with 2% bovine serum albumin (BSA) in PBS (pH 7.4) containing 0.1% Tween 20 at 25 °C. The membranes were incubated overnight in a solution containing anti-His antibody (Santa Cruz Biotechnology, sc-803) or anti-c-myc antibody (Santa Cruz Biotechnology, sc-40). The bands were visualized with ECL western blot detection kit (GE Healthcare)35 (link).
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