SDS-PAGE and immunoblotting were performed as we previously described.64 (link) Briefly, 8% SDS-acrylamide gel was used for detecting APP (1:1,000; ab12269; Abcam). 10% SDS-acrylamide gel was used for detecting BDNF (1:1,000; ab223354; Abcam), Rela (1:1,000; 8242; Cell Signaling Technology), and ApoE (1:1,000; ab183597; Abcam). 12% SDS-acrylamide gel was used for detecting CD9 (1:2,000; ab92726; Abcam), CD63 (1:1,000; ab193349; Abcam), CD81 (1:1,000; 10037; CST), neurogranin (1:1,000; ab217672; Abcam), VILIP-1 (1:1,000; ab187631; Abcam), and Aβ (1:1,000; 15126; CST). GAPDH (1:2,000; 2118; CST) and α-tubulin (1:1,000; 2144; CST) were used as the internal control. For densitometry, the ChemiDoc XRS+ Imaging System (Bio-Rad, Hercules, CA, USA) was used. Measurement of mean pixel density of each band was conducted using Quantity One software (Bio-Rad).
Ab217672
Manufactured by Abcam
Sourced in United States
Ab217672 is a lab equipment product. It is a tool used for scientific research and experimentation in a laboratory setting.
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Lab products found in correlation
3 protocols using ab217672
1
Protein Expression Analysis by SDS-PAGE and Immunoblotting
2
Protein Expression Analysis of CAMTA1 and NRGN
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The extraction of proteins were from cells and tissues of each group and the protein concentration was measured with the application of the bicinchoninic acid protein kit. After that, the protein samples were separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel, transferred onto membranes (300 mA) and blocked by 5% skimmed milk powder for 1 hour. Next, the membranes were incubated with primary antibody against CAMTA1 (ab227713, 1:1000, Abcam, Cambridge, MA, USA) and neurogranin (NRGN, ab217672, 1:1000, Abcam) (overnight, 4℃). Subsequently, the cells were incubated with horseradish peroxidase-conjugated secondary antibody (Abcam) at 37℃ for 1 h. The bands were visualized with chemiluminescent reagents. The signal intensities of the proteins were evaluated using Image J software.
3
Antibodies for Neurogranin Signaling
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The following antibodies were used in described experiments: rabbit anti-neurogranin (1:4000, Abcam, ab217672) as previously described [62, 67], rabbit anti-phosphorylated neurogranin (Ser36, 1:1000, Millipore, ABN426), rabbit anti-PSD-95 (1:1000, Abcam, ab18258), rabbit anti-phosphorylated CaMKII (Thr286, 1:5000, Cell Signaling, 12716), mouse anti-α-CaMKII (1:2000, Cell Signaling, 50049), rabbit anti-Calmodulin (1:1000, Abcam, ab45689), and mouse anti-β-actin (1:2000, Sigma, A5316).
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