Nov 21, 2023

Public workspace4% Paraformaldehye in .1M PB preparation

DOI
dx.doi.org/10.17504/protocols.io.ewov1yr6kvr2/v1
  • 1HHMI Janelia Research Campus
Monique Copeland
Open access
DOI: dx.doi.org/10.17504/protocols.io.ewov1yr6kvr2/v1
Protocol Citation: Monique Copeland 2023. 4% Paraformaldehye in .1M PB preparation. protocols.io https://dx.doi.org/10.17504/protocols.io.ewov1yr6kvr2/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 16, 2019
Last Modified: November 21, 2023
Protocol Integer ID: 26993
Funders Acknowledgement:
HHMI
Abstract
For perfusion solution.
Materials
MATERIALS
ReagentParaformaldehydeCatalog #19200
Protocol materials
ReagentSodium Hydroxide, 10.0NThomas ScientificCatalog #C871W32
Step 3
ReagentCorning™ Disposable Vacuum Filter/Storage Systems Fisher ScientificCatalog #09-761-5
Step 8
ReagentFalcon Tube (50 mL)Fischer Scientific
Step 9
ReagentSodium Phosphate Dibasic Dihydrate (USP/FCC/EP/BP), Fisher ChemicalFisher ScientificCatalog #S472-500
Step 5.1
ReagentSodium Phosphate Monobasic Anhydrous (Colorless-to-White Crystals), Fisher BioReagentFisher ScientificCatalog #BP329-500
Step 5.2
ReagentParaformaldehydeCatalog #19200
Materials, Step 4
Paraformaldehyde prep
Paraformaldehyde prep
Add Amount400 mL of MilliQ water to a Amount1000 mL beaker

Equipment
Milli-Q® Integral Water Purification System for Ultrapure Water
NAME
filter
TYPE
Millipore Sigma
BRAND
ZRXQ010WW
SKU
http://www.emdmillipore.com/US/en/product/Milli-Q-Integral-10-Water-Purification-System,MM_NF-ZRXQ010WW
LINK

Add stir bar in beaker and set on hot plate at Temperature50 °C

Add 5 drops (500ul) of 10N NaOH (from 60ml dropper)

ReagentSodium Hydroxide, 10.0NSigma AldrichCatalog #C871W32
(this makes the solution basic to help dissolve the paraformaldehyde)
Weigh Amount20 g of Paraformaldehyde and add to MilliQ water.
(Stir about Duration01:00:00 or until all parafromaldehyde is dissolved and solution is clear)

ReagentParaformaldehydeSigma AldrichCatalog #19200

Make Concentration1 Molarity (M) Phosphate buffer at PH 7.4

Prepare Concentration1 Molarity (M) Soduim Phosphate Dibasic Dihydrate (Store stock at TemperatureRoom temperature .)



ReagentSodium Phosphate Dibasic Dihydrate (USP/FCC/EP/BP), Fisher ChemicalSigma AldrichCatalog #S472-500



Prepare Concentration1 Molarity (M) Sodium Phospate Monobasic Anyhydrous (store stock at TemperatureRoom temperature )

ReagentSodium Phosphate Monobasic Anhydrous (Colorless-to-White Crystals), Fisher BioReagentSigma AldrichCatalog #BP329-500



To make Amount100 mL 1M PB at PH 7.4 add Amount19 mL Monobasic plus Amount81 mL Dibasic (store as stock at TemperatureRoom temperature )




Add Amount50 mL 1M PB PH 7.4 to paraformaldehyde solution.

Test PH and adjust solution to final desired pH using HCl or NaOH.
Top off to 500ml with MilliQ water.
Filter solution through .2um CA member filter.


ReagentCorning™ Disposable Vacuum Filter/Storage Systems Sigma AldrichCatalog #09-761-5


Lable container with '4% paraformaldehyed in .1M PB' and date. Store at Temperature4 °C for up to 1 week.
(Can also be aliqouted into
ReagentFalcon Tube (50 mL)Sigma Aldrich
and stored at Temperature-20 °C for 3 months.)