Cell Counting Kit-8 (CCK8) was obtained from Dojindo Co (Kumamoto, Japan). Puromycin was provided by Solarbio (Beijing, China). Antibodies for Tubulin (66031–1-Ig), HBO1 (13751–1-AP), Histone H4 (16047–1-AP), BCL2 (12789–1-AP), P21 (10355–1-AP), Cyclin D1 (60186–1-Ig), CDK4 (11026–1-AP), LaminB1 (66095–1-Ig), β-catenin (51067–2-AP), c-MYC (10828–1-AP) and MMP7 (10374–2-AP) were obtained from Proteintech (Wuhan, China). Antibodies for Histone H3 (#4499), acetyl-Histone H3 at Lys14 (H3K14ac, #7627), and cleaved-poly (ADP-ribose) polymerase (PARP) (#5625) were provided by Cell signaling Tech China (Shanghai, China). Antibodies for acetyl-Histone H3 at Lys23 (H3K23ac, BS70005) and acetyl-Histone H4 at Lys8 (H4K8ac, BS74016) were provided by Bioworld (Nanjing China). An acetyl-Histone H4 at Lys5 (H4K5ac, bs-10721-R) was provided by Bioss (Beijing, China). An acetyl-Histone H4 at Lys12 (H4K12ac, ab177793) was provided by Abcam (Shanghai, China). Caspase inhibitors, Z-DEVD-FMK, and Z-VAD-FMK were provided by Glpbio (Shanghai, China). Blasticidin and the HBO1 inhibitor WM-3835 were provided by MedChemExpress (Shanghai, China). Transfection reagent Lipofectamine 3000 was provided by Thermo-Fisher Invitrogen (Shanghai, China).
C myc 10828 1 ap
Manufactured by Proteintech
Sourced in United States, China
The C-Myc (10828-1-AP) is a primary antibody product offered by Proteintech. It is designed to detect the c-Myc protein, which is a transcription factor involved in the regulation of gene expression. This antibody can be used for various applications, including Western blotting, immunohistochemistry, and immunofluorescence.
Automatically generated - may contain errors
Lab products found in correlation
β actin 20536 1 ap, by Proteintech (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Lamin b1, by Proteintech (1 mentions)
Z vad fmk, by GLPBIO (1 mentions)
Histone h4, by Proteintech (1 mentions)
Mmp7 10374 2 ap, by Proteintech (1 mentions)
Puromycin, by Solarbio (1 mentions)
Phosphatase inhibitor cocktail, by Merck Group (1 mentions)
Ne pertm nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (1 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Radioimmunoprecipitation assay (ripa), by Beyotime (1 mentions)
Bcl2 associated x (bax), by Proteintech (1 mentions)
Phenylmethylsulfonyl fluoride, by Solarbio (1 mentions)
Bcl 2, by Proteintech (1 mentions)
Ubiquitin, by Cell Signaling Technology (1 mentions)
C myc, by Cell Signaling Technology (1 mentions)
Gapdh hrp 60004, by Proteintech (1 mentions)
15 protocols using c myc 10828 1 ap
1
Evaluating Cell Proliferation and Apoptosis
2
Western Blot Analysis of Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The whole cell lysates were extracted with RIPA buffer (Merck Millipore, USA) plus protease inhibitor cocktail (Sigma-Aldrich, Germany) and phosphatase inhibitor cocktail was added when phosphorylated proteins need to be detected. Nuclear and cytoplasmic proteins were extracted using NE-PERTM Nuclear and Cytoplasmic Extraction Reagents (REF78833), Thermo Scientific, USA. The proteins were separated by SDS-PAGE, transferred onto a nitrocellulose filter membrane and incubated with specific primary antibodies. The final bands were visualized with Odyssey Infrared imaging system (Li-COR Biosciences, USA) after corresponding secondary antibodies incubations. The primary antibodies are as follows: UHMK1 (#11624-1-AP), GAPDH (#60004-1-Ig), c-MYC (#10828-1-AP) and FLAG-like tag (#66008-3-Ig) were obtained from Proteintech, Wuhan, China. HA-tag (#3724), STAT3 (#9139), p-STAT3 (#9145) and Cyclin D1 (#2922) were obtained from Cell Signaling Technology, USA. For immunoprecipitation, lysis buffer without SDS (25 mmol/L Tris-HCl pH 7.4, 150 mmol/L NaCl, 1% NP-40 nonyl phenoxypolyethoxylethanol and protease inhibitor cocktail) was used for protein preparation. The procedure was performed as previously described [26 (link)].
3
Protein Expression Analysis in Cell Lysates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed in ice‐cold RIPA (Beyotime Biotechnology) containing phenylmethylsulfonyl fluoride (Solarbio). Cell protein lysates were separated by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, followed by transfer on to PVDF membranes. Membranes were then blocked with 5% nonfat milk for 1 h and incubated overnight with specific primary antibodies at 4°C. The antibodies used in the study are as follows: fructose bisphosphate 1 (FBP1; 12,842‐1‐AP, Proteintech), B‐cell lymphoma 2 (Bcl‐2; 26,593‐1‐AP, Proteintech), Bcl‐2‐associated X‐protein (Bax; 50,599‐2‐lg, Proteintech), β‐actin (20536‐1‐AP, Proteintech, China), c‐MYC (10828‐1‐AP, Proteintech) and Cyclin‐dependent kinase 4 (CDK4; 11,026‐1‐AP, Proteintech).
4
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For preparing proteins, cultured cells were lysed in radioimmune precipitation assay (RIPA) buffer plus phenylmethylsulfonyl fluoride (PMSF), protease and phosphatase inhibitors. Then the cell lysates were mixed with BCA solution and the protein concentration was determined by absorbance at 562 nm in SpectraMax M5 Multi-Mode Microplate Reader according to the instruction of the BCA protein assay kit (Beyotime, Haimen, China). Then 20 μg of protein extracts were subjected to western blot according to the standard procedure. The specific antibodies used in this study are as follows: β-Catenin (#8480), E-Cadherin (#3195), N-Cadherin (#13116), c-Myc (#9402), IgG (30000-0-AP), HA-Tag (#3724), FLAG (#14793), Ubiquitin (#3933), purchased from Cell Signaling Technology (Danvers, MA 01923, USA); c-Myc (10828-1-AP) and GAPDH (HRP-60004) purchased from Proteintech Group (Rosemont, IL 60018, USA). All antibodies were diluted to 1: 1000 in the blotting.
5
Protein Expression and Immunohistochemistry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MG132 and Liposome 2000 were obtained from Sigma (Shanghai, China). Immunohistochemical kits were obtained from ZSGB-Bio (Beijing, China). Antibodies against HECTD3 (11487-1-AP), CDK2 (10122-1-AP), CDK4 (11026-1-AP), Tubulin (11224-1-AP), HA-tag (51064-2-AP), p21 (10355-1-AP) and C-MYC (10828-1-AP) were bought through Proteintech. HECTD3 antibody (bs-15448R) was obtained from Bo Aosen Biotechnology Co., Ltd (Beijing, China) for immunohistochemistry (IHC). Antibodies Flag and Ki67 antibodies were obtained through Sigma-Aldrich (Shanghai, China).
6
Cannabinoid Pathway Regulation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CBG (purity ≥98%, B20542) was purchased from Yuanye Biotechnology Co., LTD. (Shanghai, China), Z-VAD-FMK (A1902) and cycloheximide (CHX, A8244) were purchased from ApexBio (Houston, TX, USA), Suc-Leu-Leu-Val-Tyr-AMC (ab142120) and PML-RARA antibody (AB43152) were purchased from Abcam (Cambridge, MA, USA), lithium chloride (LiCl, L9650) was purchased from Sigma-Aldrich (St. Louis, MO, USA), cleaved caspase-3 (9661 T) and β-catenin (8480S) antibodies were purchased from Cell Signalling Technology (Danvers, MA, USA), microtubule-associated protein light chain 3 (LC3, 4600-1-AP), p62 (18420-1-AP), Bax (50599-2-Ig), Bcl-2 (12789-1-AP), Pro-caspase 3 (19677-1-AP), cyclin D1 (60186-1-Ig), c-myc (10828-1-AP), C/EBP homologous protein (CHOP, 60304-1-Ig), eukaryotic initiation factor 2α (eIF2α, 66482-1-Ig), protein kinase R-like endoplasmic reticulum kinase (PERK, 24390-1-AP), and β-actin (20536-1-AP) antibodies were purchased from Proteintech (Chicago, IL, USA), and p-eIF2α (Ser51) (AF3087), p-PERK (Thr982) (DF7576), and activating transcription factor 4 (ATF4, DF6008) antibodies were purchased from Affinity Biosciences (Cincinnati, OH, USA).
7
Western Blot Analysis of Cell Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Caki-1 and SN12-PM6 cells were lysed in cold radioimmunoprecipitation assay (RIPA) lysis buffer, and the lysates were loaded onto 10–12% sodium dodecyl sulfate polyacrylamide gel for electrophoresis. Separated proteins were transferred to polyvinylidene difluoride (PVDF) membranes and probed with specific primary antibody (1:1,000 dilution) overnight at 4°C followed by incubation with 1:10,000 HRP-conjugated secondary antibody for 1 h. The blots were analyzed using enhanced chemiluminescence plus reagents and scanned with the Storm Electrophoresis Scanner (Amersham Pharmacia Biotech Inc., Piscataway, NJ, USA). The antibodies used in this study were as follows: c-Myc (10828-1-AP; Proteintech, Rosemont, IL, USA), CORO6 (17243-1-AP; Proteintech), CCND1 (60186-1-lg; Proteintech), GAPDH (60004-lg; Proteintech), and AXIN2 (ab109307; Abcam, Cambridge, UK).
8
Immortalized Cell Lines in Melanoma Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All the immortalized melanoma cell lines (A375, MV3 and SK-MEL-28), immortalized human melanocyte cell line Pig1, human immortalized epidermal cells HaCat and the human embryonic renal cell line 293FT were originally purchased from ATCC (American Type Culture Collection, Rockville, MD, USA). RAI14 (ab137118), RAI14 (ab241499) and FBXO32 (ab168372) were purchased from Abcam (Cambridge, MA, USA); α-Tubulin (2146), c-MYC (#9402S) and HA-Tag (3724S) were purchased from Cell Signaling Technology (Boston, MA, USA). P21 (10355-1-AP), CDK4 (66950-1-Ig), CCND1 (60186-1-Ig), GAPDH (60004-1-Ig), FBXO32 (67172-1-Ig), RAI14 (17507-1-AP) and c-MYC (10828-1-AP) were purchased from Proteintech (Wuhan, China). MG132 (#S2619) was obtained from Selleck Chemicals (Shanghai, China). Cycloheximide (CHX, #C7698) was purchased from Sigma-Aldrich (Shanghai, China). ECL reagents was obtained from Beyotime (#P0018, Shanghai, China). RIPA lysis buffer was purchased from Beyotime (Shanghai, China). Certified Fetal Bovine Serum (FBS) was purchased from VivaCell (VivaCell, Shanghai, China). Propidium iodide (PI) was purchased from BD Biosciences. Dulbecco’s modified Eagle’s medium (DMEM) was purchased from BD Becton, Dickinson and Company.
9
Protein Expression Analysis by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Twenty micrograms of the total protein was separated using Biofuraw™ Precast Bis-Tris Gel (180-8008H, Tanon) and then transferred onto PVDF membranes. After being incubated with 5% skim milk in TBST for 1 h, the membranes were incubated with specific primary antibodies including c-Myc (10828-1-AP, Proteintech), loricrin (LOR) (55439-1-AP, Proteintech), transglutaminase 1 (TGM1) (12912-3-AP, Proteintech), O-GlcNAc (MA1-072, Thermo Scientific) or β-actin (66009-1-Ig, Proteintech) overnight at a dilution of 1:1000 at 4°C. Then, appropriate secondary antibodies were chosen for incubation for 1 h at room temperature. Finally, immunoreactive protein bands were imaged using a horseradish peroxidase substrate for enhanced chemiluminescence (WBKLS0050, Millipore). Gray intensity analysis of immunoblots was evaluated by the Image-Pro Plus software.
10
Protein Expression Analysis in HaCaT Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein was extracted from HaCaT cells and measured with the BCA assay. Proteins (20 μg) were separated by Biofuraw™ Precast Bis-Tris Gel (180-8008H, Tanon, Shanghai, China) and then transferred to nitrocellulose membranes. After blocking with 5% skimmed milk for 1 h, the membranes were then incubated overnight at 4°C with the following primary antibodies at the dilution of 1:1000: c-Myc (10828-1-AP, Proteintech, Rosemont, IL, USA), transglutaminase 1 (TGM1,12912-3-AP, Proteintech), loricrin (LOR, 55439-1-AP, Proteintech), keratin 1(K1,16848-1-AP, Proteintech), β-catenin (51067-2-AP, Proteintech), H3 (A2348, ABclonal, Woburn, MA, USA), S100A6 (10245-1-AP, Proteintech) and β-actin (66009-1-Ig, Proteintech). The next day, the membranes were incubated with the HRP-conjugated secondary antibody (1:5000) for 1 h at room temperature. Finally, western blot bands were visualised using electrochemical luminescence. The band density analysis was performed using Image‐Pro Plus.
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!