Antibodies to NF-κB p65 (no. 8242S), beta-tubulin (no. 2128S), GAPDH (no. 5174S), phospho-MAPK p38 (T180+Y182; no. 4511S), total MAPK p38 (no. 8690S), and total SAPK/JNK (no. 9252T) were obtained from Cell Signaling Technology. Antibodies to phospho-JNK (T183+Y185; no. 6254), phospho-FOS (S374; no. 81485), total c-FOS (no. 166940), phospho-JUN (S63; no. 822), and total c-JUN (no. 74543) were obtained from Santa Cruz Biotechnology. Goat anti-rabbit and anti-mouse IgG-horseradish peroxidase (HRP) secondary antibodies (no. 31460 and no. 31430) were obtained from Thermo Fisher Scientific. DAPI nuclear stain was obtained from Cell Signaling, and goat anti-rabbit IgG AlexaFluor488 was obtained from Molecular Probes (Life Technologies).
Total sapk jnk
Manufactured by Cell Signaling Technology
Sourced in United States
Total SAPK/JNK is a lab equipment product that measures the total amount of SAPK/JNK protein in a sample, regardless of its activation state. It provides a quantitative assessment of the SAPK/JNK protein levels in a given biological system.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay, by Thermo Fisher Scientific (2 mentions)
Total c jun, by Santa Cruz Biotechnology (1 mentions)
Alexa fluor 488 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Phloretin, by Merck Group (1 mentions)
Phospho sapk jnk, by Cell Signaling Technology (1 mentions)
Sb203580, by Merck Group (1 mentions)
Total erk1 2, by Cell Signaling Technology (1 mentions)
Anti β actin antibody, by Merck Group (1 mentions)
Cell culture medium and supplements, by Thermo Fisher Scientific (1 mentions)
Ara a, by Merck Group (1 mentions)
Phospho p38α mapk, by Cell Signaling Technology (1 mentions)
Total p38α mapk, by Cell Signaling Technology (1 mentions)
Phospho erk1 2, by Cell Signaling Technology (1 mentions)
Pd98059, by Merck Group (1 mentions)
Sp600125, by Merck Group (1 mentions)
Phospho sapk jnk thr183 tyr185, by Cell Signaling Technology (1 mentions)
Cell lysis buffer, by Cell Signaling Technology (1 mentions)
P19arf, by Abcam (1 mentions)
Total p53, by Abcam (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
4 protocols using total sapk jnk
1
Immunoblotting Analysis of Cell Signaling Pathways
2
AMPK Signaling Pathway Regulation
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Cell culture medium and supplements were purchased from GIBCO-BRL (Rockville, MD, USA). Phloretin, the AMPK inhibitor ara-A, the MEK inhibitor PD98059, the JNK inhibitor SP600125, the p38 inhibitor SB203580, and the anti-β actin antibody were purchased from Sigma–Aldrich (St. Louis, MO, USA). Antibodies against phospho-AMPKα (Thr172), total AMPKα, phospho-ERK1/2, total-ERK1/2, phospho-SAPK/JNK, total-SAPK/JNK, phospho-p38α MAPK, and total-p38α MAPK were purchased from Cell Signaling Technology (Beverly, MA, USA).
3
Western Blot Analysis of Cell Signaling
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Cells were incubated in cell lysis buffer (no. 9803; Cell Signaling Technology, Danvers, MA) followed by centrifugation at 13,000 rpm for 10 minutes. Supernatants were collected, and protein concentrations were determined using the Bradford Assay (Bio-Rad, Hercules, CA). Protein lysates were electrophoresed on Tris-glycine SDS polyacrylamide gels and transferred onto Immobilon-P membrane. Incubation with antibodies was performed according to Cell Signaling Technology-recommended procedures and proteins were visualized using ECL (Pierce). p16Ink4A, p19ARF, p21, Total-p53, cdc2 were from Abcam. Phospho-p53(Ser15), Cyclin B1, Phospho-SAPK/JNK(Thr183/Tyr185), Total-SAPK/JNK and β-Actin were from Cell Signaling Technology.
4
Western Blot Analysis of Stress Kinases
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After RIPA buffer lysis (Cell Signaling, Leiden, The Netherlands), protein concentration was measured with BCA Protein assay (Thermo Fisher Scientific). 30 μg protein extracts were resolved by 10% SDS PAGE, transferred to nitrocellulose membranes and probed with antibodies against: phospho-stress-activated protein kinase/c-Jun NH(2)-terminal kinase (SAPK/JNK) (Thr183/Tyr185, clone G9 MAB #9255), total SAPK/JNK (#9252), phospho-p38 Mitogen-Activated Protein Kinase (MAPK) (Thr180/Tyr182, clone 28B10 MAB #9216), total p38 MAPK (#9212) (all from Cell Signaling), p75NTR (clone 8211 MAB5264) (EMD Millipore), GAPDH (clone G9, MAB #32233) (Santa Cruz Biotechnology), and tubulin (Sigma Aldrich). Blots were developed by ECL system (Amersham Biosciences) according to the manufacturer’s protocol.
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