For western blot analysis, the proteins separated on SDS-PAGE gels were transferred onto a polyvinylidene fluoride membrane with a BioRad electroblotting apparatus as described [40 (link)]. The signals on the blots were detected with anti-p44/42 MAP kinase antibody (Cell Signaling Technology, MA) or anti-GFP antibody (Thermo Fisher Scientific, USA) under the ECL Supersignal system (Pierce, Rockford, IL).
Anti p44 42 map kinase antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Anti-p44/42 MAP kinase antibody is a laboratory tool used to detect the expression and phosphorylation of the p44/42 mitogen-activated protein (MAP) kinases, also known as ERK1/2. This antibody recognizes the unphosphorylated and phosphorylated forms of these kinases, which are important signaling molecules involved in a variety of cellular processes.
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Lab products found in correlation
Electroblotting apparatus, by Bio-Rad (1 mentions)
Anti gfp antibody, by Thermo Fisher Scientific (1 mentions)
Phospho sapk jnk, by Cell Signaling Technology (1 mentions)
Phospho syk, by Cell Signaling Technology (1 mentions)
Phospho gab2, by Cell Signaling Technology (1 mentions)
Phospho p44 42 mapk erk1 2, by Cell Signaling Technology (1 mentions)
Phospho plcγ2, by Cell Signaling Technology (1 mentions)
Phospho specific antibody, by Cell Signaling Technology (1 mentions)
Phospho p38 map kinase, by Cell Signaling Technology (1 mentions)
Phospho src family, by Cell Signaling Technology (1 mentions)
Alexa 488 conjugated anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Alexa 594 conjugated anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Phospho nf kb, by Cell Signaling Technology (1 mentions)
Phospho akt, by Cell Signaling Technology (1 mentions)
Anti phospho egfr tyr1068 antibody, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase hrp linked anti rabbit igg, by Cell Signaling Technology (1 mentions)
Anti phospho erk1 2 antibody, by Cell Signaling Technology (1 mentions)
Anti egfr antibody, by Cell Signaling Technology (1 mentions)
Biotin peac5 maleimide bpm, by Dojindo Laboratories (1 mentions)
Anti mouse igg 7076 secondary antibody, by Cell Signaling Technology (1 mentions)
4 protocols using anti p44 42 map kinase antibody
1
Western Blot Immunodetection Procedure
2
Phospho-Specific Antibody Sourcing for Cell Signaling
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Antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), with the exception of anti-p44/42 MAP Kinase antibody and the following phospho-specific antibodies which were obtained from Cell Signaling Technologies (Danvers, MA, USA): Phospho Syk (Tyr525/526); Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204); Phospho-SAPK/JNK (Thr183/Tyr185); Phospho-p38 MAP Kinase (Thr180/Tyr182); Phospho-Src Family (Tyr416); Phospho-NFkB (Ser536); Phospho-Gab2 (Tyr452); Phospho-PLCγ2 (Tyr1217); Phospho-Akt (Ser473). The anti-human C12orf4 antibody and all reagents were obtained from Sigma-Aldrich (St Louis, MO, USA). Antiphosphotyrosine mAb 4G10 was purchased from Upstate Biotechnology (Millipore, MA, USA). Alexa 488 conjugated anti-mouse IgG and Alexa 594 conjugated anti-rabbit IgG antibodies were purchased from Jackson ImmunoResearch laboratories (West Grove, PA, USA).
3
Quantification of Cellular Redox Signaling
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Epidermal growth factor (EGF), p-nitrophenyl phosphate (pNPP), and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). 1,2-NQ (95% purity determined by HPLC), 1,4-NQ (100% purity determined by GC), and 1,4-BQ (99.1% purity determined by iodometric titration) were purchased from Tokyo Chemical Industry (Tokyo, Japan).
PD153035 and anti-PTP1B antibody (# PH01) were acquired from Calbiochem (San Diego, CA, USA). Biotin-(PEAC) 5 -maleimide (BPM) and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) were obtained from Dojindo Laboratories (Kumamoto, Japan). The anti-EGFR antibody (#2232), anti-phospho EGFR Tyr1068 antibody (#2234), anti-p44/42 MAP kinase antibody [anti-extracellular signal-regulated kinase 1/2 (ERK1/2), #9102], anti-phospho ERK1/2 antibody (#9101), horseradish peroxidase (HRP)-linked anti-rabbit IgG (#7074), and anti-mouse IgG (#7076) secondary antibody were procured from Cell Signaling Technology (Beverly, MA, USA). Fetal bovine serum, GlutaMAX-I, tris(2-carboxyethyl) phosphine hydrochloride (TCEP) and Ni-IDA ProBond were from Corning (Woodland, CA, USA), Gibco (Grand Island, NY, USA), Hampton Research (Aliso Viejo, CA, USA) and Invitrogen (Carlsbad, CA, USA), respectively. The anti-1,4-NQ antibody was prepared as previously reported (Hirose et al., 2012) . All other reagents used were of the highest purity available.
PD153035 and anti-PTP1B antibody (# PH01) were acquired from Calbiochem (San Diego, CA, USA). Biotin-(PEAC) 5 -maleimide (BPM) and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) were obtained from Dojindo Laboratories (Kumamoto, Japan). The anti-EGFR antibody (#2232), anti-phospho EGFR Tyr1068 antibody (#2234), anti-p44/42 MAP kinase antibody [anti-extracellular signal-regulated kinase 1/2 (ERK1/2), #9102], anti-phospho ERK1/2 antibody (#9101), horseradish peroxidase (HRP)-linked anti-rabbit IgG (#7074), and anti-mouse IgG (#7076) secondary antibody were procured from Cell Signaling Technology (Beverly, MA, USA). Fetal bovine serum, GlutaMAX-I, tris(2-carboxyethyl) phosphine hydrochloride (TCEP) and Ni-IDA ProBond were from Corning (Woodland, CA, USA), Gibco (Grand Island, NY, USA), Hampton Research (Aliso Viejo, CA, USA) and Invitrogen (Carlsbad, CA, USA), respectively. The anti-1,4-NQ antibody was prepared as previously reported (Hirose et al., 2012) . All other reagents used were of the highest purity available.
4
Protein Phosphorylation Analysis in Fungal Mutants
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The mutants and Guy11 strains were cultured in liquid CM for 2 days and then the total proteins were isolated from vegetative hyphae as described by Bruno et al. (2004 (link)). For protein phosphorylation analysis, the proteinase inhibitor cocktail (cOmplete; Sigma‐Aldrich) was added. The intensity of the signal corresponding to phosphorylated MoMps1 was detected by binding of an antiphospho‐p44/42MAP kinase antibody (Cell Signaling Technology), and the anti‐p44/42 MAP kinase antibody (Cell Signaling Technology) was used as control. For green fluorescent protein (GFP)‐tagged protein detection, samples were analysed by 8% SDS‐PAGE followed by western blotting with the anti‐GFP antibody (Abmart) and the anti‐mouse antibody (LI‐COR, IRDye), followed by detection using the ODYSSEY infrared imaging system (application software v. 2.1).
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