The following antibodies were purchased from Santa Cruz: monoclonal anti-HA, rabbit polyclonal anti-HA and polyclonal anti-Cdc20. Rabbit polyclonal antibodies to Histone H3, Cdc27 and PHistone-H2AX (Ser139) antibodies were purchased from Cell Signalling Technologies. The following antibodies were purchased from Sigma-Aldrich: monoclonal anti-g-tubulin, polyclonal α-tubulin, HRP-conjugated goat anti-mouse, HRP-conjugated goat anti-rabbit. A human anti-centromere antibody was purchased from Europa Bioproducts Ltd. Monoclonal antibodies to PCNA, Mad2B and c-myc were purchased from BD Transduction Laboratories. The following antibodies were purchased from Invitrogen: Alexa Fluor 488 goat anti-mouse IgG, Alexa Fluor 488 goat anti-rabbit IgG, Alexa Fluor 594 rabbit anti-mouse IgG and Alexa Fluor 594 goat anti-human IgG.
C myc
Manufactured by BD
Sourced in United States
The C-Myc is a laboratory equipment product offered by BD. It is designed to measure the expression levels of the c-Myc protein, which is a transcription factor involved in various cellular processes. The core function of the C-Myc is to provide accurate and reliable quantification of the c-Myc protein in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (2 mentions)
Rabbit polyclonal anti ha, by Santa Cruz Biotechnology (2 mentions)
Alexa fluor 594 rabbit anti mouse igg, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 594 goat anti human igg, by Thermo Fisher Scientific (2 mentions)
Polyclonal anti cdc20, by Santa Cruz Biotechnology (2 mentions)
Polyclonal α tubulin, by Merck Group (2 mentions)
Hrp conjugated goat anti mouse, by Merck Group (2 mentions)
Hrp conjugated goat anti rabbit, by Merck Group (2 mentions)
Alexa fluor 488 goat anti mouse igg, by Thermo Fisher Scientific (2 mentions)
E cadherin, by BD (2 mentions)
Vimentin, by BD (2 mentions)
Nanog, by Abcam (2 mentions)
Protease inhibitor cocktail, by Roche (2 mentions)
Phistone h2ax ser139, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase, by Merck Group (1 mentions)
Monoclonal anti γ tubulin, by Merck Group (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Tra 1 81, by BD (1 mentions)
Ssea1, by BD (1 mentions)
Tra 1 60, by BD (1 mentions)
17 protocols using c myc
1
Identification of Mitotic Regulators
2
Comprehensive Antibody Acquisition Protocol
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The following antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA): monoclonal anti-HA, rabbit polyclonal anti-HA and polyclonal anti-Cdc20. Rabbit polyclonal antibodies to Chk1, PChk1 (Ser317), Cdc27, PHistone-H2AX (Ser139), Histone H3 antibodies and mouse monoclonal FLAG antibody were purchased from Cell Signaling Technology (Beverly, MA, USA). The following antibodies were purchased from Sigma-Aldrich (St Louis, MO, USA): monoclonal anti-γ-tubulin, polyclonal α-tubulin, horseradish peroxidase (HRP)-conjugated goat anti-mouse, HRP-conjugated goat anti-rabbit. A human anti-centromere antibody was purchased from Europa Bioproducts Ltd (Cambridge, UK). Monoclonal antibodies to Mad2B, Mad2A, PCNA and c-myc were purchased from BD Transduction Laboratories (Heidelberg, Germany). The following antibodies were purchased from Invitrogen (Carlsbad, CA, USA): Alexa Fluor 488 goat anti-mouse IgG, Alexa Fluor 488 goat anti-rabbit IgG, Alexa Fluor 594 rabbit anti-mouse IgG and Alexa Fluor 594 goat anti-human IgG.
3
Induced Pluripotent Stem Cell Characterization
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Goat iPS cells were stained according to the AKP staining kit instructions (SiDanSai- Cat. No.: 1101–050). We washed the cultured cells 24 h and 21 d post-transfection with PBS for 2–3 times. We subsequently performed indirect immunofluorescence by following the method of Zhang et al. [11 (link)]. The dilution ratio of anti-rabbit antibody was 1:1000, and the dilution ratio of FITC-labeled goat anti-rabbit secondary antibody was 1:1000. We added DAPI at a ratio of 1:100, and performed nuclear staining for 10 min. We observed and photographed the cells using a fluorescence microscope (Olympus- Cat. No.: IX51). The detail antibody information were provided as below: OCT4 (Abcam- Cat. No.: ab19857, dilution ratio 1:500), SOX2 (Abcam- Cat. No.:ab97959, dilution ratio 1:500), KLF4 (Abcam- Cat. No.: ab72543, dilution ratio 1:500), C-MYC (BD Biosciences- Cat. No.: 551101, dilution ratio 1:500), CDX2 (BD Biosciences- Cat. No.: 560171, dilution ratio 1:500), REX (Abcam- Cat. No.: ab50828, dilution ratio 1:500), SSEA-1(BD Biosciences- Cat. No.: 561585, dilution ratio 1:500), TRA-1-60 (BD Biosciences- Cat. No.: 560884, dilution ratio 1:500), TRA-1-81 (BD Biosciences- Cat. No.: 560072, dilution ratio 1:500).
4
Western Blot Analysis of Cell Signaling
5
Western Blot Analysis of EMT and Stemness Markers
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Cell lysates were harvested in ice-cold modified radioimmune precipitation assay buffer containing a protease inhibitor cocktailTM (Roche). Lysates normalized for amount protein were separated on 10% SDS-polyacrylamide gels and electrophoretically transferred to nitrocellulose. The blots were incubated with primary antibodies to E-CADHERIN, VIMENTIN, FIBRONECTIN, β-CATENIN (BD), c-MYC, FOXM1, TFIIB, MYD88, γ-CATENIN, SNAIL1, SLUG, TWIST, hCTR1 (Santa Cruz), BMI1, CD44, NANOG, ALDH-1, OCT-4, NOTCH-1, SFRP5 (Abcam), ZO-1 (Invitrogen), phospho-β-CATENIN, N-CADHERIN (Cell Signaling), or SOX-2 (Sigma). Immunocomplexes were detected with horseradish peroxidase-conjugated IgG and visualized via enhanced chemiluminescence (ECL detection kit, Amersham).
6
Staining and Detection of Stemness Markers
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For staining with OCT-4, Sox-2 and c-Myc, cells were fixed and permeabilized with Cytofix-Cytoperm kit (BD Pharmingen, San Diego, Ca, USA) according to the manufacturer's instructions. Cells were then incubated with monoclonal mouse IgG2b anti-human OCT-3/4, Sox-2 and c-Myc (Santa Cruz Biotechnology, Santa Cruz, Ca, USA) at 4°C for 30 minutes, washed twice with PBS and incubated with corresponding secondary Abs [phycoerythrin (PE)-conjugated polyclonal goat anti-mouse] at 4°C for 30 minutes. hEpCAM-FITC (Biosource, CA, USA), biotin-conjugated anti-human CD133 as primary Abs (Miltenyi Biotec, CA, USA) and streptavidin-FITC (BD Pharmingen, USA) as a secondary Abs were used.
7
Antibodies for Cell Signaling Pathways
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This has been performed as previously described [12 (link)]. Antibodies directed against Vimentin (RV202), Twist1, N-cadherin and interleukin-6 (IL-6) were purchased from Abcam (Cambridge, MA); Akt, phospho-Akt (193H12), Erk1/2, phospho-Erk1/2 (THR202/TYR204), E-cadherin (24E10) and MCP-1 from Cell Signaling (Danvers, MA); c-Myc from BD Biosciences (San Jose, CA); Cyclin D1 (HD11), ERα (F-10) and glyceraldehydes-3-phosphate dehydrogenase (GAPDH, FL-335), were purchased from Santa Cruz (Santa Cruz, CA).
8
Protein Extraction and Western Blotting
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Cells were solubilized in lysis buffer comprising phosphate-buffered saline solution containing 1× cell lysis buffer (Cell Signaling Technology, Danvers, MA, USA), 1× protease inhibitor cocktail (Roche, Indianapolis, IN, USA), and 1× phosphatase inhibitor cocktail (Roche). The mixtures were incubated for 30 min on ice. The lysates were, then, centrifuged for 10 min at a speed of 13,000 rpm at 4°C. Total protein concentrations were determined using the Bio-Rad Protein Assay Kit (Bio-Rad, Hercules, CA, USA) according to the manufacturer’s instructions. Total proteins (20 μg) were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Bio-Rad) and transferred to polyvinylidene-fluoride membranes (0.45 μm, Millipore, Bedford, MA, USA), and subsequently probed with first and second antibodies. The following antibodies were used: α-tubulin (Sigma-Aldrich, St Louis, MO, USA), p21WAF1/CIP1, HIF1α (BD Biosciences, San Jose, CA, USA), c-Myc, PP2Aα, p-4EBP1, 4EBP-1, p-S6 ribosomal proteinSer235/Ser236, S6 ribosomal protein, p-p38 MAPK, p38 MAPK, and horseradish peroxidase-linked anti-mouse and anti-rabbit IgG (all from Cell Signaling Technology).
9
Protein Signaling Pathway Analysis
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Cells were cultured without drug, with memantine, AraC (60 nM for Jurkat, 250 nM for Molm-13), and memantine plus AraC. Cytoplasmic and nuclear protein extracts were prepared as described [37 (link)]. Antibodies used in Western blots were: p-AKT(S473), AKT, pS6(S240/244), p-ERK1/2(T202/Y204), ERK1/2, pJNK1/2(Thr183/Tyr185), c-JUN (60A8), human Caspase-9, active Caspase-3 (5A1E, only detects cleaved active Caspase-3), Caspase-8 (all Cell Signaling, Leiden, The Netherlands), c-MYC (9E10, BD Pharmingen), β-Actin (AC-74, Sigma-Aldrich), Lamin B (sc-6217, Santa Cruz Biotechnology Europe). Primary antibodies were detected with species-specific secondary antibodies (Dianova, Hamburg, Germany) and chemiluminescence. Nitrocellulose membranes were reprobed for several proteins.
10
Western Blot Analysis of Pluripotency Markers
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The whole lysate of GEFs from post-transfection day 1, 6, 9, 12, 15, 18, and 21 was extracted by following the protocol recommended by the protein extraction kit manufacturer. Western blots were performed by following the methods reported [17 (link)]. The detail antibody information were provided as below: Oct 4 (Abcam- Cat. No.: ab19857, dilution ratio 1:1000), Sox 2 (Abcam- Cat. No.:ab97959, dilution ratio 1:1000), Klf 4 (Abcam- Cat. No.: ab72543, dilution ratio 1:1000), C-Myc (BD Biosciences- Cat. No.: 551101, dilution ratio 1:1000), Nanog (Abcam- Cat. No.: ab21624, dilution ratio 1:1000), β-actin (Abcam- Cat. No.: ab8226, dilution ratio 1:1000), goat anti-mouse IgM [FITC] labeled (Abcam - Cat. No.: ab8227, dilution ratio 1:1000).
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