Cells were lyzed using 0.20% NP40, 50 mM Tris pH 7.5, 5% Glycerol, 1.5 mM MgCl2, 100 mM NaCl lysis buffer containing Phosphatase Inhibitor Cocktail 2 (Sigma, P5726) and cOmplete Protease Inhibitor Cocktail (Roche, 11873580001). Lysates were resolved by SDS-PAGE and incubated with primary antibodies. Antibodies used were against actin (A5441), CAMKK2 (HPA017389) (both Sigma), ERK1/2 (Sigma, M5670) and phospho-AKT (Ser473)(#9271), AKT (#9272), ALK(#3633), phospho-p90RSK (Ser380)(#12032), RSK1/2/3 (#9355), RSK1(#9333), RSK2 (#5528), phospho-RPS6 (Ser235/236)(#4858), RPS6 (#2217), phospho-ERK1/2 (Thr202/Tyr204)(#4267), phospho-FAK1 (Tyr397)(#8556), FAK1 (#13009), phospho-IGF1R (Tyr1131)(#3021), IGF1R (#9750), cleaved Caspase 3 (#9661), PARP-1 (#9542), AMPK1 (#2795), pAMPKα (Thr172)(#2535), FER (#4268), phospho-YB1 (Ser102)(#2900), and YB1 (#4202) were from Cell Signaling. Secondary antibodies were HRP-conjugated α-rabbit or α-mouse (GE Healthcare).
Phospho yb1
Manufactured by Cell Signaling Technology
Sourced in United States
Phospho-YB1 is a lab equipment product from Cell Signaling Technology. It is an antibody that specifically detects the phosphorylated form of the YB1 protein.
Automatically generated - may contain errors
Lab products found in correlation
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Phosphatase inhibitor cocktail 2, by Merck Group (1 mentions)
Erk1 2, by Merck Group (1 mentions)
Phospho akt, by Merck Group (1 mentions)
Phospho igf 1r, by Cell Signaling Technology (1 mentions)
P ampkα, by Cell Signaling Technology (1 mentions)
Rsk1 2 3, by Cell Signaling Technology (1 mentions)
Complete protease inhibitor cocktail, by Roche (1 mentions)
Phospho fak1, by Cell Signaling Technology (1 mentions)
Hrp conjugated α rabbit, by GE Healthcare (1 mentions)
Phospho p90rsk, by Cell Signaling Technology (1 mentions)
Igf 1r, by Cell Signaling Technology (1 mentions)
Parp1, by Cell Signaling Technology (1 mentions)
Phospho erk1 2, by Cell Signaling Technology (1 mentions)
Phospho rps6, by Cell Signaling Technology (1 mentions)
Cleaved parp, by Cell Signaling Technology (1 mentions)
Phospho akt, by Cell Signaling Technology (1 mentions)
Imagequant las 4000 mini, by GE Healthcare (1 mentions)
Phospho erk, by Cell Signaling Technology (1 mentions)
Cyclin d1, by Abcam (1 mentions)
3 protocols using phospho yb1
1
Comprehensive Cell Lysis and Immunoblotting
2
Protein Expression Analysis in Cultured Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RIPA lysis buffer was used to lyse the cultured cells (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and a BCA Protein Assay Kit (Pierce Biotechnology, Rockford, IL, USA) was applied to determine protein concentrations. Antibodies to YB-1 (1:1000, Cell Signaling Technology, Danvers, MA, USA), GAPDH (1:1000, Cell Signaling Technology, Danvers, MA, USA), Cleaved PARP (1:1000, Cell Signaling Technology, Danvers, MA, USA), cyclinD1 (1:1000, Epitomics, Burlingame, CA, USA), ERK (1:1000, Cell Signaling Technology, Danvers, MA, USA), phospho-ERK (1:1000, Cell Signaling Technology, Danvers, MA, USA), phospho-YB-1 (1:1000, Cell Signaling Technology, Danvers, MA, USA), AKT (1:1000, Cell Signaling Technology, Danvers, MA, USA), phospho-AKT (1:1000, Cell Signaling Technology, Danvers, MA, USA), mTOR (1:1000, Cell Signaling Technology, Danvers, MA, USA) and phospho-mTOR (1:1000, Cell Signaling Technology, Danvers, MA, USA) were used. The anti-mouse/rabbit IgG secondary antibodies (1:2000, Cell Signaling Technology, Danvers, MA, USA) were used for 1 h. The bound proteins were visualized using electrochemiluminescence (GE Healthcare, Tokyo, Japan) and were quantitated using a ImageQuant LAS 4000 Mini (GE Healthcare, Tokyo, Japan).
3
Protein Analysis of Cell Lysates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were harvested and whole‐cell lysates were prepared using lysis buffer (50 mM HEPES [pH 7.5], 150 mM NaCl, 5 mM EDTA, 1% NP‐40, and 0.65% CHAPS containing 1 mM PMSF and a protease inhibitor cocktail). Sample preparation and Western blot analysis were carried out as previously described18 using antibodies against the following proteins: Hsp70, AR (N‐20), β‐actin, c‐Jun, and Twist (Santa Cruz Biotechnology, Dallas, TX, USA); Hsp90, PSA, YB‐1, phospho‐YB‐1, and phospho‐Foxo3a (Cell Signaling Technology, Danvers, MA, USA); AR‐V7 (Precision Antibody, Columbia, MD, USA); UBE2C (BostonBiochem, Cambridge, MA, USA); FKBP5, CREB, and Sp1 (Gene Tex, CA, USA); laminB1 (Abcam, Cambridge, UK); Foxo3a (Upstate, Temecula, CA, USA); α‐tubulin (Calbiochem, San Diego, CA, USA); and HRP‐conjugated secondary antibodies (GE Healthcare, Little Chalfont, UK).
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!