For immunoblot analysis, cells were washed twice with PBS and lysed in lysis buffer (1% NP-40, 120 mM NaCl, 40 mM Tris-HCl (pH7.4), 1.5 mM sodium orthovanadate, 50 mM sodium fluoride, 10 mM sodium pyrophosphate, and protease inhibitor cocktail (Roche)), as described in (Cai et al. 2020 (link); Lee et al. 2020 (link)). Primary antibodies against phospho-Akt (9272), Akt (4056), phospho-ERK (9101), ERK (4696) (Cell Signaling), α-tubulin (T5169; Sigma), and horseradish peroxidase-conjugated secondary antibodies were used. IPMK antibody was generated with a peptide targeting mouse IPMK amino acids 295-311 (SKAYSRHRKLYAKKHQS) from Covance.
Phospho erk 9101
Manufactured by Cell Signaling Technology
Sourced in United Kingdom
Phospho-ERK (9101) is a laboratory reagent produced by Cell Signaling Technology. It is a primary antibody that specifically detects ERK1/2 proteins phosphorylated at Thr202/Tyr204 and Thr185/Tyr187. This antibody can be used to detect and quantify the activation of the ERK1/2 signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Erk 4696, by Cell Signaling Technology (1 mentions)
α tubulin, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Ab32445, by Abcam (1 mentions)
Ab182126, by Abcam (1 mentions)
Ab133485, by Abcam (1 mentions)
Phorbol 12 myristate 13 acetate tpa, by Merck Group (1 mentions)
Erk 4695, by Cell Signaling Technology (1 mentions)
Anti β actin a1978 antibody, by Merck Group (1 mentions)
Recombinant human hgf 100 39, by Thermo Fisher Scientific (1 mentions)
Nf κb p65 sc 8008, by Santa Cruz Biotechnology (1 mentions)
Anti iκbα sc 371, by Santa Cruz Biotechnology (1 mentions)
3 protocols using phospho erk 9101
1
Immunoblotting Protocol for Protein Analysis
2
Immunoblotting for Kinase Signaling
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Western blotting was followed, was described elsewhere [12 (link)]. Polyclonal rabbit antibody for Fli-1 (ab133485), PKCδ (ab182126), Phospho- PKCδ (ab133456), MEK (ab178876), Phospho-MEK (ab96379), BAD (ab32445), and Phospho-BAD (ab129192) were obtained from Abcam (Abcam, Cambridge, UK); ERK (#4695) and phospho-ERK (#9101) from Cell Signalling Technology (CST, Danvers, MA01923, USA), β-actin (20536–1-AP) and GAPDH (13937–1-AP), from Proto-Technology (Protein-Tech, Bucuresti, Romania). Antibody dilution according to the manufacturer instructions.
The inhibitor of MEK (U0126) [#S1102] were obtained from Sellectchem (Sellectchem, Houston, USA) and PKC agonist Phorbol 12-myristate 13-acetate (TPA) from Sigma, (Sigma, St. Louis, MO, USA). These compounds were dissolved in DMSO and added to the cells at indicated concentration, as described in the results section.
The inhibitor of MEK (U0126) [#S1102] were obtained from Sellectchem (Sellectchem, Houston, USA) and PKC agonist Phorbol 12-myristate 13-acetate (TPA) from Sigma, (Sigma, St. Louis, MO, USA). These compounds were dissolved in DMSO and added to the cells at indicated concentration, as described in the results section.
3
Antibody Sourcing for Malaria Research
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Antibodies against PKCζ (9372), MET (4560), AKT (9272s), phospho-AKT (9271), ERK1/2 (9102), and phospho-ERK (9101) were obtained from Cell Signaling. CXCR4 (60042–1-Ig), NGFR (55014–1-AP), KRBP (10386–1-AP), SULF2 (12260–1-AP), and dihydrofolate reductase (15194–1-AP) were obtained from Proteintech. Antibodies against CD160 (ab202845), C1orf114 (ab121933), and CTRP4 (ab36871) were obtained from Abcam. Anti-IκBα (sc-371) and NF-κB p65 (sc-8008) were obtained from Santa Cruz. Anti–β-actin antibody (A1978) was obtained from Sigma-Aldrich. Antibodies against Plasmodium UIS4 (LS-C204260/53255) and Plasmodium HSP70 (LS-C109068/25028) were obtained from LSBio. Anti-P. falciparum CSP (MRA-183A) was obtained from MR4. Polyclonal anti-UIS3 was raised in a rabbit against its recombinant protein, which was produced by the pET system (Clontech) in Escherichia coli. Anti-AMA1 was kindly provided by Prakash Srinivasan (Johns Hopkins Malaria Research Institute, Baltimore, MD). Recombinant human HGF (100-39) was obtained from Peprotech.
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