The only way to solubilize many antigens for immunoprecipitation is by denaturation. This cell lysis protocol is ideally suited for this purpose to release proteins from complex structures or reveal antibody epitopes hidden within native proteins. Short linear epitopes may not be accessible to antibodies within the native tertiary and quaternary protein structures, but they become exposed upon the unraveling of proteins, exposing their secondary structure. Antibodies otherwise not suitable for the immunoprecipitation of proteins prepared under nondenaturing conditions are now able to bind these antigens of interest in cell lysates prepared under denaturing conditions. These antibodies may also work well for immunoblotting purposes when the protein target is completely denatured. Harvested cells in this protocol are washed in tris-buffered saline (TBS) before lysis in 2% sodium dodecyl sulfate (SDS)-containing Lysis buffer for 10 min at 100°C. The resulting sample is diluted 20-fold in TBS to reduce the SDS concentration to ≤0.1% before the addition of an antibody for immunoprecipitation. Addition of 2% bovine serum albumin (BSA) or 0.1% Nonidet P-40 to the TBS before an immunoprecipitation, respectively, ensures either removal of SDS from the target protein or retaining denatured proteins in solution.

Differential detergent fractionation of cells is a rapid method for extraction of cytoplasmic and nuclear proteins in preparation of an immunoprecipitation. This method can be applied for use of adherent or suspension cells and can significantly reduce nonspecific background in an immunoprecipitation by separation of cellular compartments into individual fractions. The lysis of cells by differential detergents permits the rapid extraction of proteins from the cytoplasm (digitonin), the cytoplasmic membranes, and organelles (Triton X-100), and nucleoplasm (Tween/DOC), facilitated through the use of distinct extraction buffers. Cytoplasmic and nuclear matrix proteins as well as DNA are left behind during the detergent-based extraction.

This rapid and efficient method to prepare highly purified recombinant adeno-associated viruses (rAAVs) is based on binding of negatively charged rAAV capsids to an anion-exchange resin that is pH dependent.

This is a simple method for rapid preparation of recombinant adeno-associated virus (rAAV) stocks, which can be used for in vivo gene delivery. The purity of these vectors is considerably lower than that obtained by either CsCl gradient centrifugation or by combination of iodixanol gradient ultracentrifugation followed by column chromatography.

The most commonly used method for production of recombinant adeno-associated virus (rAAVs) in research laboratories is by transient triple transfection of 293 cells with AAV cis and trans plasmids and an adenovirus helper plasmid. This protocol describes the processes required to prepare the transfected cell suspension for virus purification.

The Bradford assay is a quick and fairly sensitive method for measuring the concentrations of proteins. It is based on the shift in absorbance maximum of Coomassie Brilliant Blue G-250 dye from 465 to 595 nm following binding to denatured proteins in solution.

FSI Briefs No 1, April 2020. Regulatory policy responses should seek to support economic activity while preserving the financial system's soundness and ensuring transparency. The recommendation for banks to make full use of capital and liquidity buffers should go hand in hand with restrictions on dividends and bonuses and clarity concerning the process for rebuilding them. Flexibility in loan classification criteria for prudential and accounting purposes should be complemented with sufficient disclosure on the criteria banks use to assess creditworthiness. The publication of detailed guidance on the application of expected loss provisioning rules, combined with sensible transitional arrangements, may constitute a balanced approach to mitigating the unintended effects of the new accounting standards.

FSI Briefs No 2, April 2020. Guidance issued by financial sector authorities in response to the Covid-19 crisis seems to suggest that international efforts to come up with operational resilience standards should take into account at least the following elements: Critical/essential employees: identifying the critical functions and employees that support important business services, as well as ensuring employees' safety and that they can safely resume their duties (remotely, if necessary); IT infrastructure: ensuring that IT infrastructure can support a sharp increase in usage over an extended period and taking steps to safeguard information security; Third-party service providers: ensuring that external service providers and/or critical suppliers are taking adequate measures and are sufficiently prepared for a scenario in which there will be heavy reliance on their services; Cyber resilience: remaining vigilant in order to identify and protect vulnerable systems, and detect, respond and recover from cyber attacks..

Using a cross-country sample of bank-dependent public fi rms we study the international spillovers of a change in banking regulation on corporate borrowing. For identi cation we examine how US rms' liabilities vis-a-vis banks, non-bank lenders and bond markets evolve after an increase in capital requirements implemented by the European Banking Authority (EBA) in 2011. We find that US firms experience a reduction in credit lines but not in term loans from EU banks.

FSI Briefs No 3, April 2020. In response to the 2007-09 Great Financial Crisis (GFC), accounting standard setters introduced a new methodology to value loans based on expected credit losses (ECL). The previous approach, based on incurred losses, was viewed as procyclical and inconsistent with prudential objectives. In the wake of the Covid-19 pandemic, several prudential authorities and the Basel Committee on Banking Supervision (BCBS), introduced a series of measures to clarify how banks should consider various public and private debt relief programmes in their ECL estimates and in their calculation of regulatory capital. These measures are intended to incentivise banks to continue supporting the real economy, while reducing pressure on banks' ECL provisions, earnings and regulatory capital. Supervisory initiatives that provide capital relief should be augmented by severe constraints on the payment of dividends, bonuses and share buybacks. These joint actions will simultaneously expand banks' lending capacity and enhance their ability to absorb losses. Prudential authorities face difficult trade-offs as they confront the most severe economic crisis in modern times. Encouraging the use of flexibility in applicable accounting standards, while preserving market trust and transparency in the reported financial statements of banks, will be key in fostering both economic and financial stability.

BIS Bulletin No 8, April 2020. Information on local labour markets and Google searches can be used to construct a measure of the vulnerability of employment in different regions of the United States to the Covid-19 shock. Regional exposure to Covid-19 varies significantly, ranging from a low of 2% to a high of 98% of total local employment. We test for the usefulness of the Covid-19 exposure measure by showing that areas with higher exposure report more Google search queries related to the pandemic and unemployment benefits.

BIS Bulletin No 9, April 2020. By allowing banks to run down some of their buffers, policymakers are sending a strong signal about their resolve to lessen the economic fallout from the pandemic. Such prudential measures complement the main policy levers: monetary and fiscal instruments. To avoid a reduction in credit to the real economy, authorities need to ensure that banks have the capacity and willingness to make use of the flexibility afforded by the buffer release. Payout restrictions on banks and risk-sharing between banks and the public sector will be key. For banks to continue playing a positive role in the supply of funding during the recovery, they should maintain usable buffers for a long period, as losses from a severe recession will take time to materialise.

Recent literature has highlighted that international trade is mostly priced in a few key vehicle currencies, and is increasingly dominated by intermediate goods and global value chains (GVCs). Taking these features into account, this paper reexamines the business cycle dynamics of international trade and its relationship with monetary policy and exchange rates.

The National Book Festival Presents series was created to provide a book festival experience to lovers of the event on a year-round basis. Because these programs can no longer be held at the Library, we are offering a virtual multipart series, with authors who have written books about widespread diseases and the worldwide response to them.

A prototype deployed in San Francisco Bay imagines the underside of a floating building as an upside-down artificial reef

The graphic designer is receiving a Lifetime Achievement Award from Cooper Hewitt for her recognizable computer icons, typefaces and graphics

British product designer Lucy Hughes has invented a biodegradable plastic made from fish offcuts

The Wisconsin-born architect's buildings helped turn the city he once called an 'inglorious mantrap' into the center of the world

BIS Press Release - Easing trade tensions lift sentiment: BIS Quarterly Review, 8 December 2019

Press release: CPMI report sets out considerations for developers of wholesale digital tokens, 12 December 2019

Press release: Basel Committee invites comments on the design of a prudential treatment for crypto-assets, 12 December 2019.

Markets Committee calls for wider adoption of global code of conduct for foreign exchange markets (Press release, 30 January 2020)

No global real estate market despite higher price synchronisation and growing role of international investors, central banks find (Press release, 18 February 2020)

Press release "Governors and Heads of Supervision announce deferral of Basel III implementation to increase operational capacity of banks and supervisors to respond to Covid-19", 27 March 2020

BCBS Press release "Basel Committee and IOSCO announce deferral of final implementation phases of the margin requirements for non-centrally cleared derivatives", 3 April 2020

Press release "Saudi G20 Presidency and the Bank for International Settlements (BIS) Innovation Hub invite global innovators to solve RegTech and SupTech challenges", 27 April 2020

Tissue engineering has a unique set of tools and technologies for developing preventive strategies, diagnostics, and treatments that can play an important role during the ongoing COVID-19 pandemic.

A protein shredder that occurs in cell membranes of brain cells apparently also indirectly regulates the fat metabolism.

The number of people evaluated for signs of stroke at U.S. hospitals has dropped by nearly 40% during the COVID-19 pandemic, according to a study led by researchers from Washington University School of Medicine in St. Louis who analyzed stroke evaluations at more than 800 hospitals across 49 states and the District of Columbia.

How the PleaseNotSanders movement achieved what NeverTrump never could.

Will a nationalist president be undone by his underreaction to a foreign threat?

It’s the coronavirus election now.

The purpose of suffering may be mysterious, but the search for meaning is obligatory.

The last time Charlene Chiddenton saw her father in person was on March 7 — visitor restrictions at long-term care homes during the pandemic had kept them apart — and less than two months later, she was saying goodbye to her father by video call.

When the province eventually conducts its review on the response to COVID-19, the president of the Nova Scotia Nurses’ Union can think of one thing in particular that needs to be discussed.

Classified released a music video for his song, Good News, this week featuring a pack of kids hanging out with seniors at a continuing care home in Enfield, N.S. It was filmed before the COVID-19 outbreak in the province.

Graduation is on hold for Dalhousie University medical student Manveer Bal, so he's decided to spend his newfound free time volunteering at the epicentre of the COVID-19 outbreak in Nova Scotia.

A Dalhousie University scientist and a NSCAD textile professor are teaming up to create a new fabric. It could be used to locally manufacture personal protective equipment like surgical masks. As the CBC's Colleen Jones reports, they have received a COVID-19 grant to study the idea.

Nova Scotia students and teachers will not be returning to the classroom this year. At-home learning will continue until June 5, when the province's school year will end.

What would have been Cape Breton University's largest convocation to date is being replaced by an online offering this year because of COVID-19. Some graduating students say it just won't be the same as a traditional ceremony.

The latest death was at the Northwood long-term care home in Halifax.

Quebec's public health director, Dr. Horacio Arruda, says grandparents under 70 years old are allowed to see and even care for their grandchildren. But the recommendation comes with cautions.

The COVID-19 crisis has wreaked havoc on Quebec's economy, with the province recording its highest unemployment rate since at least 1976, when the data was first tracked.

The provincially appointed trustee of the English Montreal School Board says the Quebec government made the “right decision” when it postponed the reopening of Montreal-area elementary schools to May 25.

Here is what you should know as Quebec loosens travel restrictions in some regions.

The current deconfinement plan in the Montreal area could lead to a rapid increase in deaths in the city, according to findings by Quebec's public health institute.