Primary antibodies against ADAM8, EGFR, CCL2, and Akt (panAkt and pAkt S473) were obtained from Proteintech; p-EGFR, ERK1/2 (panERK and pERK1/2 Tyr 202/204) from Cell Signaling Technology (Danvers, MA, USA); HB-EGF from ABclonal technology (Woburn, MA, USA). Iba-1, GFAP, and CD206 were purchased from Abcam (Cambridge, UK). Secondary antibodies were listed in the Supplementary Data Table S1 together with primary antibodies. Temozolomide was purchased from Selleckchem (Houston, TX, USA). As an EGFR inhibitor, we used Erlotinib (Selleckchem).
Pakt s473
Manufactured by Proteintech
Sourced in United States
The PAkt S473 is a laboratory equipment product that measures the phosphorylation level of the Ser473 residue of the Akt protein. It provides a quantitative assessment of this specific post-translational modification without further interpretation or extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Temozolomide, by Selleck Chemicals (1 mentions)
Erlotinib, by Selleck Chemicals (1 mentions)
Iba 1, by Abcam (1 mentions)
Panakt, by Proteintech (1 mentions)
Hb egf, by ABclonal (1 mentions)
Glial fibrillary acidic protein (gfap), by Abcam (1 mentions)
Cd206, by Abcam (1 mentions)
Minute total protein extraction kit, by Invent Biotechnologies (1 mentions)
Anti p erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)
Aldh1a1, by Proteintech (1 mentions)
Erk1 2, by Wanlei (1 mentions)
Oct3 4, by Wanlei (1 mentions)
β actin, by Beyotime (1 mentions)
Gapdh, by Beyotime (1 mentions)
Ab6789, by Abcam (1 mentions)
Ab6721, by Abcam (1 mentions)
Erbb2, by Santa Cruz Biotechnology (1 mentions)
Ipvh00010, by Merck Group (1 mentions)
Gapdh, by Abcam (1 mentions)
Ripa buffer, by Solarbio (1 mentions)
3 protocols using pakt s473
1
EGFR Inhibition in Glioblastoma
2
Protein Extraction and Western Blotting Protocol
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Detailed procedure was described previously.54 (link) Protein in fresh tissues was extracted using Minute Total Protein Extraction Kit (Invent). Antibodies against the following proteins were used: YY1 (1:1,000, Wanleibio), OCT3/4 (1:1,000, Wanleibio), ALDH1A1 (1:1,000, Proteintech), p-AKT-s473 (1:1,000, Proteintech), AKT (1:1,000, Wanleibio), anti-p-ERK1/2 (Thr202/Tyr204; 1:1,000, Cell Signaling Technology), ERK1/2 (1:1,000, Wanleibio), GAPDH (glyceraldehyde-3-phosphate dehydrogenase 1:2,000, Beyotime), and β-actin (1:5,000, Beyotime). Protein expression levels were quantified by density analysis using Quantity One Software and normalized to β-actin or GAPDH.
3
Western Blot Analysis of Cardiomyocyte Proteins
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Total proteins from H9c2 cardiomyocytes were homogenized or lysed with RIPA buffer (Solarbio, R0010) and quanti ed by the Bicinchoninic Acid (BCA) protein assay kit (Thermo Scienti c, 23223). The amount of 30 µg protein lysate were separated by SDS-PAGE gels and then transferred to polyvinylidene uoride (PVDF) membranes (Merck Millipore, IPVH00010). After blocking blocked with 5% BSA in Tris-buffered saline containing 0.1% (v/v) Tween 20 (TBST) for 2 h at room temperature, the membrane were incubated with appropriate primary antibodies overnight at 4°C. Then, membranes were incubated with either HRP-goat-anti-mouse (Abcam, ab6789) or HRP-goat-anti-rabbit (Abcam, ab6721), to bind the primary antibodies for 2 h at room temperature. Next, immunoreactive bands were visualized by ChemiDoc MP Imaging System using WesternBright ECL HRP Substrate (Advansta, K-12045-D50) and quantitatively analyzed with ImageJ software.
The primary antibodies used in this study are as follows: Bax (CST, #2772); Cleaved Caspase-3 (C-CAS-3) (CST, #9661); HO-1 (CST, #43966); ErbB2 (Santa Cruz, sc-7301); p-AKT (S473) (Proteintech, 66444-1-Ig); AKT (Proteintech, 10176-2-AP); Nrf2 (Proteintech, 16396-1-AP). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Abcam, ab9485) was used as the internal reference to normalized protein expression levels.
The primary antibodies used in this study are as follows: Bax (CST, #2772); Cleaved Caspase-3 (C-CAS-3) (CST, #9661); HO-1 (CST, #43966); ErbB2 (Santa Cruz, sc-7301); p-AKT (S473) (Proteintech, 66444-1-Ig); AKT (Proteintech, 10176-2-AP); Nrf2 (Proteintech, 16396-1-AP). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Abcam, ab9485) was used as the internal reference to normalized protein expression levels.
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