Sodium fluoride and orthovanadate, phenylmethylsulfonyl fluoride, aprotinin and leupeptin were purchased from Sigma (Saint-Louis, MO, USA). Anti-phospho-mTor, anti-mTor, anti-phospho-P70S6kinase (thr389), anti-P70S6kinase, anti-phospho-S6 Ribosomal Protein (ser235-236), anti-S6 Ribosomal Protein, anti-LC3-b, anti-phospho-DRP1 (ser616) and HRP conjugated anti-rabbit antibodies were from Cell Signaling Technology (Beverly, MA, USA). Anti-Hsp60, anti-Hsp90 and anti-DRP1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-Smac antibody was purchased from RnD systems (Minneapolis, MN, USA). HRP-conjugated anti-mouse and anti-goat antibodies were from Dakopatts (Glostrup, Denmark).
Anti p70 s6 kinase
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-p70 S6 kinase is a primary antibody that specifically recognizes the p70 S6 kinase protein. The p70 S6 kinase is a serine/threonine kinase that plays a key role in the regulation of cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Anti phospho p70 s6 kinase, by Cell Signaling Technology (12 mentions)
Anti akt, by Cell Signaling Technology (11 mentions)
Anti mtor, by Cell Signaling Technology (10 mentions)
Anti 4e bp1, by Cell Signaling Technology (8 mentions)
Anti phospho p70 s6 kinase thr389, by Cell Signaling Technology (7 mentions)
Anti phospho akt, by Cell Signaling Technology (5 mentions)
Anti caspase 3, by Cell Signaling Technology (5 mentions)
Anti phospho mtor, by Cell Signaling Technology (4 mentions)
Anti phospho 4ebp1, by Cell Signaling Technology (4 mentions)
Anti gapdh, by Cell Signaling Technology (4 mentions)
Anti phospho ampkα, by Cell Signaling Technology (4 mentions)
Anti ampkα, by Cell Signaling Technology (4 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (4 mentions)
Anti β actin, by Merck Group (3 mentions)
Anti parp, by Cell Signaling Technology (3 mentions)
Anti phospho akt ser473, by Cell Signaling Technology (3 mentions)
Anti β actin, by Cell Signaling Technology (3 mentions)
Anti bcl 2, by Cell Signaling Technology (3 mentions)
Anti s6 ribosomal protein, by Cell Signaling Technology (2 mentions)
Anti lc3b, by Cell Signaling Technology (2 mentions)
34 protocols using anti p70 s6 kinase
1
Immunoblotting Analysis of Mitochondrial Proteins
2
Western Blot Analysis of Metabolic Enzymes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell lysates were prepared and subjected to WB analysis as performed previously51 (link),52 (link). The following commercial antibodies were used: anti-β-actin (Sigma A1978), anti-PKM2 (Cell Signaling Technology 3198S), anti-PKM1 (Cell Signaling Technology 7067), anti-PKM (Abcam AB118499), anti-human specific LDHA (Cell Signaling Technology 2012S), anti-LDHB (Abcam AB75167), anti-human and rabbit LDHA (Abcam AB135396), anti-p70 S6 kinase (Cell Signaling Technology 2708T), anti-calreticulin (Cell Signaling Technology 12238T), anti-golgin (Cell Signaling Technology 13192), anti-COX IV (Proteintech 11242-1-AP), anti-GPT2 (Santa Cruz sc-398383), anti-PHGDH (Sigma HPA021241), and anti-PC (Santa Cruz sc-271493), horseradish peroxidase-conjugated secondary antibodies anti-mouse (Fisher PI31430) and anti-rabbit (Fisher PI31460). All the primary antibodies were used at a 1:500 dilution in 5% non-fat milk in TBST. Secondary antibodies were used at a 1:3000 dilution in 5% non-fat milk in TBST.
3
Western Blot Analysis of Liver Protein Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Liver homogenates and cells were lysed in ice-cold RIPA buffer (50 mM Tris HCl (pH 7.4), 250 mM NaCl, 1% Nonidet P-40, and protease inhibitor cocktail). The protein samples (50 µg) were separated by 4%–12% sodium dodecyl sulfate polyacrylamide gel electrophoresis and then transferred onto PVDF membranes. After blocking with 5% BSA or 5% non-fat dry milk, the membranes were incubated overnight with the following primary antibodies: anti-p-mTOR, anti-mTOR, anti-p-p70s6kinase, anti-p70s6kinase, anti-p-4EBP-1, anti-4EBP-1, anti-CHOP, anti-p-eIF2α, and anti-eIF2α (Cell Signaling Technology, Boston, MA, USA), anti-ubiquitin, anti-GRP78, anti-p-PERK, anti-PERK, anti-Bax, anti-lamp-1, anti-Tom20, anti-cathepsin B, and β-actin (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-p62 (SQSTM1; MBL International, Woburn, MA, USA), and anti-LC3II (Novus Biologicals, Littleton, CO, USA). Immunoreactive bands were visualized using the ECL Western blotting protocol (Bio-Rad). Finally, the membranes were exposed to imaging film (Kodak BioFlexEcono Scientific Supplies, Citrus Heights, CA, USA), after which the film was developed using a Kodak X-OMAT 1000A Processor. Densitometric analysis of the bands was conducted using the Image J software (NIH, Bethesda, MD, USA).
4
Investigating Protein Regulation via Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analysis was done using the whole-cell extracts prepared in RIPA buffer. The blot was probed with antibodies of a polyclonal antibody raised against human p53 K139 acetylated peptide, anti-human p53 (DO-1, Santa Cruz), anti-Phospho-p70 S6 kinase (Thr389; Cell Signaling Technologies), anti-p70 S6 kinase (Cell Signaling Technologies 49D7), anti-phospho-4E-BP1 (Ser65; Cell Signaling Technologies 9451), anti-4E-BP1 (Cell Signaling Technologies 9644), anti-DDIT4 (Proteintech 10638-1-AP), and anti-mouse p53 (Novocastra CM5). The levels of β-actin (Sigma-Aldrich A3853) were used as the protein loading control.
5
Molecular Pathway Regulation in Cancer
6
Western Blot Analysis of Metabolic Enzymes
7
Evaluating Signaling Pathways in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H2O2 (30%) and crystal violet was purchased from Sigma (St. Louis, MO, USA). MTT reagent was purchased from Duchefa Biochemie (Haarlem, Netherlands). anti-pY701-STAT1, anti-STAT1, anti-pY705-STAT3, anti-STAT3, pY694-STAT5, anti-STAT5, anti-pT202/Y204-p44/42 MAPK (Erk1/2), anti-p44/42 MAPK (Erk1/2), anti-pT183/Y185-SAPK/JNK and anti-SAPK/JNK, anti-pT180/Y182-p38, anti-p38, anti-pS473-Akt, anti-Akt, anti-PT389-p70S6 kinase, anti- p70S6 kinase, anti-pS133-CREB, anti-CREB were purchased from Cell Signaling Technology (Danvers, MA, USA). anti-Synaptophysin, anti-PSD95, anti-pT205-Tau, anti-pS262-Tau, and anti-Tau were purchased from ABclonal (Wuhan, China). anti-α-Tubulin was purchased from Abbkine (Wuhan, China). All other chemicals were obtained from Sigma-Aldrich (St. Louis, MO, USA).
8
Comprehensive Western Blot Analysis of Akt Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A western blot analysis was performed with anti‐phosphorylated Akt at Ser473 (Cat. No. 4060, Cell Signaling Technology, Danvers, MA, USA), anti‐phosphorylated Akt at Thr308 (Cat. No. 4056, Cell Signaling Technology), anti‐Akt (Cat. No. 4691, Cell Signaling Technology), anti‐phosphorylated p70 S6 kinase at Thr389 (Cat. No. 9234, Cell Signaling Technology), anti‐p70 S6 kinase (Cat. No. 2708, Cell Signaling Technology), anti‐irisin (Cat. No. G‐067‐17, Phoenix Pharmaceuticals), anti‐phosphorylated focal adhesion kinase (FAK) at Tyr397 (Cat. No. 3283, Cell Signaling Technology), anti‐FAK (Cat. No. 13009, Cell Signaling Technology), and anti‐glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) antibodies (Cat. No. 5174, Cell Signaling Technology) as described previously.10
9
Mitomycin C and Rapamycin Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mitomycin C was from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Rapamycin and protease inhibitor cocktail were obtained from Sigma-Aldrich (St Louis, MO, USA). S6K1WT-HA, S6K1E389DeltaCT-HA and S6K1F5A were purchased from Addgene (Cambridge, MA, USA). Anti-HA, anti-phospho-Akt, anti-Akt, anti-CEA, anti-phospho-JNK, anti-JNK, anti-Bak, anti-caspase 8, anti-caspase 9, anti-caspase 3, anti-phospho-p70 S6 kinase (Thr389), anti-p70 S6 kinase, anti-phospho-Bad (Ser136), anti-Bad, anti-COX-IV and anti-PARP antibody were from Cell Signaling (Danvers, MA, USA). Anti-actin antibody was from Sigma-Aldrich. Anti-cytochrome c antibody was from BD PharMingen (San Jose, CA, USA). Anti-Ki67 was purchased from Dako (Carpinteria, CA, USA).
10
Metformin Effects on Autophagy Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells treated or not with metformin (10mM), were lysed in 70μL of lysis buffer (50mM Tris HCl pH 7.5, 0.1% Triton, 5mM EDTA complemented by protease (Chemicon Millipore) and phosphatase (Sigma-Aldrich) inhibitors. Western blots were performed as previously described using monoclonal rabbit antibodies [20 (link), 22 (link)], anti-LC3b (1/1000, Cell Signaling), anti-Beclin 1 (1/1000, Cell Signaling), anti-p62 (1/1000 Abcam), anti-phospho (T172) AMPK (1/1000, Cell Signaling), anti-AMPK (1/1000, Cell Signaling) anti-phospho (S79) ACC (1/1000, Cell Signaling), anti-ACC (1/1000, Cell Signaling), anti-phospho (S2448) mTOR (1/1000, Cell Signaling), anti-mTOR (1/1000, Cell Signaling), anti-phospho (T389) p70S6 Kinase (1/1000, Cell Signaling), anti-p70S6 Kinase (1/1000, Cell Signaling), anti-phospho (T37/46) 4EBP1 (1/1000, Cell Signaling), anti-phospho (S473) AKT (1/1000, Cell Signaling), anti-phospho (T308) AKT (1/1000, Cell Signaling), anti-AKT (1/1000, Cell Signaling), anti-HIF-1α (1/1000, Cayman Chemical), anti-Redd1/DDIT4 (1/1000, Abcam and Proteintech) and were normalized using a rabbit polyclonal antibody anti-β-tubulin (1/1000, Cell Signaling). Gel quantification was performed using ImageJ (Windows 1.47, Research Services Branch, NIH).
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!