Application
Western blot
Sample
Mouse Tissue lysate - whole (Mouse Brain)
Specification
Mouse Brain
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Dilution
1/1000
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 3% milk in Tris buffered saline +Tween20
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti mouse
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/3000
Detection
Detection method
ECL
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 200, 70 kDa
Additional data
PubMed ID
17682049: View
Additional Notes
Forebrains from the postnatal 1 d (n = 4), 1 wk (n = 2), and 10 wk (n = 2) MMP-deficient or WT mice were homogenized with buffer containing 0.32 M sucrose, 10 mM Hepes, pH 7.4, 2 mM EDTA, 50 mM NaF, 1 mM Na2VO4, and 1x protease cocktail inhibitors (Roche), using a glass-teflon homogenizer. The homogenates were then centrifuged at 1,000 g, and the supernatants were centrifuged at 50,000 rpm to separate the membrane fractions from the soluble fractions. The membrane fractions were suspended in lysis buffer (1% Triton X-100, 50 mM Hepes, pH 7.4, 2 mM EDTA, and protease/phosphatase inhibitors). For Western blotting, 20 µg of protein from each sample was suspended in Laemmli sample buffer.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
DR. Li Tian
Verified customer
投稿 Sep 21 2007