For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

We use cookies to make our site as useful as possible.

Our Cookie Policy explains how you can opt-out of the cookies we use.

If you continue without changing your cookie settings, we'll assume you’re happy with this.

Continue Continue

United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Diagnostic & Therapeutic Solutions
    Custom solutions & partnerships

    Custom antibody development and commercial partnerships to advance your diagnostic and therapeutic discovery.

    Create custom solutions with us

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

Supporting our customers and employees during the COVID-19 pandemic. Read more

Products by research area:
  • Cancer
  • Cardiovascular
  • Cell Biology
  • Epigenetics & Nuclear Signaling
  • Developmental Biology
  • Immunology
  • Metabolism
  • Microbiology
  • Neuroscience
  • Signal Transduction
  • Stem Cells

Brain Synaptosomal Preparation

By Dr. Taylor Kimberly *

  1. Homogenize brain tissue. Add 5 ml of 0.32 M sucrose. Homogenize with 12 strokes of a 19x84mm tissue grinder (Potter Elvehjem, plastic coated) at 800 rpm.
  2. Centrifuge to remove large debris. Centrifuge for 10 min at 1000 g at RT.  Remove 100 ul for synaptophysin analysis.
  3. Layer supernatant onto sucrose and centrifuge again.
  4. Carefully layer the supernatant onto 4 ml of 1.2 M sucrose in a SW41 centrifuge tube (Beckman). 
  5. Spin at 160,000 g for 15 min (= 33,000 rpm with SW41 rotor).
  6. Carefully remove synaptosome layer. The synaptosomes are at the interface between the 1.2 M and 0.32 M sucrose layers. It is a slightly cloudy thin layer. Mitochondria and lysosomes pellet to the bottom. 
  7. Remove 100 ul for synaptophysin analysis.
  8. Dilute synaptosomes with 0.32 M sucrose. Add 4 ml of 0.32 M sucrose to synaptosomes, mix, and then carefully layer onto 4 ml of 0.8 M sucrose in a fresh centrifuge tube.
  9. Centrifuge to pellet synaptosomes.
  10. Spin at 160,000 g for 15 min. The pellet is enriched in synaptosomes.
  11. Discard the supernatant and resuspend in 1 ml of 1X STE. 
  12. Remove 50 ul for electron microscopy analysis.
  13. Lyse synaptosomes for immunoprecipitation analysis. Add 110 ul of 10% NP-40 STEN lysis (- BSA) and nutate at 4°C for 20 min. Protein normalize samples, remove sample for synaptophysin analysis then continue IP according to standard protocols.

Buffers

2.55 M sucrose (unbuffered)

  • Heat on a stirring block to dissolve, then bring up to 1 L.
  • Determine concentration using a refractometer and adjust to 2.55 M sucrose (refractive index is 1.4558 for 87.28% sucrose (w/v) at 2.5498 M)

10% NP-40 STEN lysis buffer (-BSA)

  • 5 ml 2x STEN
  • 1 ml NP-40
  • 0.1 ml 100X PI
  • 0.1 ml 100x Pefabloc
  • 3.8 ml ddH2O
  • Store at -20°C.

Note:

amyloid beta in brain -

if value = 20 pM

then 20 pmole/Liter = 20 pmole/1000g

so x 12 for dilution = 240 pmole/1000g = 0.24 pmol/g


Harvard Medical School/Brigham and Women's Hospital

Get resources and offers direct to your inbox Sign up
A-Z by research area
  • Cancer
  • Cardiovascular
  • Cell biology
  • Developmental biology
  • Epigenetics & Nuclear signaling
  • Immunology
  • Metabolism
  • Microbiology
  • Neuroscience
  • Signal transduction
  • Stem cells
A-Z by product type
  • Primary antibodies
  • Secondary antibodies
  • Biochemicals
  • Isotype controls
  • Flow cytometry multi-color selector
  • Kits
  • Loading controls
  • Lysates
  • Peptides
  • Proteins
  • Slides
  • Tags and cell markers
  • Tools & Reagents
Help & support
  • Support
  • Make an Inquiry
  • Protocols & troubleshooting
  • Placing an order
  • RabMAb products
  • Biochemical product FAQs
  • Training
  • Browse by Target
Company
  • Corporate site
  • Investor relations
  • Company news
  • Careers
  • About us
  • Blog
Events
  • Tradeshows
  • Conferences
International websites
  • abcam.cn
  • abcam.co.jp

Join with us

  • LinkedIn
  • facebook
  • Twitter
  • YouTube
  • Terms of sale
  • Website terms of use
  • Cookie policy
  • Privacy policy
  • Legal
  • Modern slavery statement
© 1998-2021 Abcam plc. All rights reserved.