Two to three days after transfection, HEK cells were harvested and homogenized in lysis buffer (10 mM Tris‐HCl 6.8, 2.5% SDS, 2 mM EDTA), containing protease inhibitor cocktail (#P8340, Sigma), phosphatase inhibitor cocktail 2 (#P5726, Sigma) and phosphatase inhibitor cocktail 3 (#P0044, Sigma). Protein concentration in the samples was determined using the BCA protein assay kit (Pierce) and then lysate concentrations were adjusted to 1 mg/ml. Western blots were probed with primary antibodies against CAMK2G (C‐18; raised against the 478–495 C‐terminal peptide, 1:1000, sc‐1541, Santa Cruz; validated in this study by overexpression experiments), CAMK2A (6G9, 1:40.000, Abcam; validated in Elgersma et al. (2002 )), CAMK2B (CB‐β1, 1:10.000, Invitrogen; validated in van Woerden et al. (2009 )), Actin (MAB1501R, 1:20.000, Chemicon; validated in Antibodypedia (https://Antibodypedia.com ), Ph‐Thr286/Thr287 (auto‐phosphorylated CAMK2 antibody; #06‐881; 1:1000; Upstate Cell Signaling Solutions; validated in Elgersma et al. (2002 )) and RFP (#600401379, 1:2000, Rockland, validated in this study by overexpression experiments), and secondary antibodies (goat anti‐mouse (#926‐32210), goat anti‐rabbit (#926‐68021), and donkey anti‐goat (#926‐68074), all 1:15.000, LI‐COR). Blots were quantified using LI‐COR Odyssey Scanner and Odyssey 3.0 software.
Cb β1
Manufactured by Thermo Fisher Scientific
CB‐β1 is a lab equipment product manufactured by Thermo Fisher Scientific. It is a core component designed for specific laboratory applications. The detailed description and specifications of this product are not available at this time.
Automatically generated - may contain errors
Lab products found in correlation
Odyssey scanner, by LI COR (2 mentions)
Actin mab1501r, by Merck Group (1 mentions)
Sc 1541, by Santa Cruz Biotechnology (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Phosphatase inhibitor cocktail 2, by Merck Group (1 mentions)
Phosphatase inhibitor cocktail 3, by Merck Group (1 mentions)
Mab1501r, by Merck Group (1 mentions)
2 protocols using cb β1
1
Quantification of CAMK2 Expression and Phosphorylation
2
Brain CAMK2 Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were anaesthetized using isoflurane and sacrificed by decapitation. Brain samples were taken out quickly and stored in liquid nitrogen. Upon protein determination lysates were first prepared and brain samples were homogenized in lysis buffer (10 mM Tris-HCl 6.8, 2.5% SDS, 2 mM EDTA). Protein concentration in the samples was determined and then lysate concentrations were adjusted to 1 mg/ml. Western blots were probed with primary antibodies against either CAMK2A (6G9, 1:40.000, Abcam), CAMK2B (CB-β1, 1:10.000, Invitrogen), Actin (MAB1501R, 1:20.000, Chemicon) or Ph-T286/T287 (autophosphorylated α CAMK2A and CAMK2B antibody; #06–881; 1:5000; Upstate Cell Signaling Solutions) and secondary antibodies (goat anti-mouse and/or goat anti-rabbit, both 1:3000, AffiniPure #115-007-003 and #111-007-003). Blots were stained with Enhanced ChemoLuminescence (ECL) (#32106, Pierce) or stained and quantified using LI-COR Odyssey Scanner and Odyssey 3.0 software. Quantification of western blot in ECL was done using ImageJ.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!