For most immunoblots developed with chemiluminescence or with fluorochrome-based detection systems, it is possible to remove the primary and secondary antibodies from the membrane without affecting the bound antigen. This allows you to reuse the membrane for detection of another protein antigen. The blots developed with chromogenic substrates can also be stripped of antibodies and reprobed, but the bands detected in the first round of immunoblotting will remain unaffected. Stripping and reprobing of the membrane are particularly useful when the amount of sample is limited or when it is important to accurately compare the signal between two different protein antigens in the same sample. Examples of such experiments include determining the levels of a protein antigen in a series of samples relative to the loading control and comparison of the phosphorylated form to the total levels of the protein in the sample.

Successful modification of the bacterial artificial chromosome (BAC) after two-step BAC engineering is confirmed in two separate polymerase chain reactions (PCRs). The first reaction (5' co-integrate PCR) uses a forward 5' co-integrate primer (a sequence located upstream of the 5' end of the A-box) and a reverse 3' primer on the vector (175PA+50AT) or within the reporter sequence or mutated region as appropriate. The second reaction (3' co-integrate PCR) uses a forward 5' primer on the recA gene (RecA1300S) and a reverse 3' co-integrate primer (a sequence located downstream from the 3' end of the B-box). Those colonies shown to be positive in PCR analysis are further tested for sensitivity to UV light. After the resolution, colonies that have lost the excised recombination vector including sacB and recA genes become UV light sensitive.

Bacterial artificial chromosome (BAC) clones are rendered electrocompetent and transformed with the recombinant shuttle vector, pLD53SCAB/AB-box. Cointegrates are selected by growth on chloramphenicol and ampicillin to ensure recombination of the shuttle vector into the BAC.

Plasmid DNA is prepared from the recombinant shuttle vector pLD53.SCAB/A-B created by cloning of the A and B homology arms for two-step bacterial artificial chromosome (BAC) engineering. To confirm that the A-box and B-box arms have been successfully incorporated into pLD53.SCAB, the pattern of enzyme digestion of the modified plasmid is compared with that of the unmodified pLD53.SCAB. Once the shuttle vector is shown to carry the proper sequences, it is ready for transfer into the BAC host.

This protocol describes the preparation of the shuttle vector before its introduction into bacterial artificial chromosome (BAC) host cells for BAC two-step engineering. The homology arm sequences, prepared previously, are introduced by ligation into the digested shuttle vector DNA to provide sites for recombination within the BAC clone. Crude lysates of individual bacterial transformants serve as templates in polymerase chain reaction (PCR) analysis to confirm the presence of the homology arms in the recombinant shuttle vector.

The 700-bp A homology arm (A-box) and the 700-bp B homology arm (B-box) are amplified by polymerase chain reaction (PCR) using purified bacterial artificial chromosome (BAC) DNA as template for two-step BAC engineering. The resulting A-box PCR product contains an AscI site at its 5' end (the 5' primer incorporates an AscI site, and the 3' primer does not incorporate any restriction sites). The B-box PCR product contains an XmaI site at its 3' end (the 5' primer does not incorporate any restriction sites, and the 3' primer incorporates an XmaI site). The amplification products are then digested with the appropriate restriction endonucleases to render them suitable for cloning into the shuttle vector.

In two-step bacterial artificial chromosome (BAC) engineering, a single plasmid is introduced into the BAC-carrying cell lines. The shuttle vector pLD53.SCAB (or pLD53.SCAEB) carries the recA gene and the R6K origin, which requires the protein to replicate. PIR2 cells, expressing , are typically used for the amplification of the vector and maintain about 15 copies/cell of the donor vector, which is relatively stable in this host.

CPMI report: Payment aspects of financial inclusion in the fintech era, April 2020

BIS Bulletin No 7, April 2020. Past epidemics had long-lasting effects on economies through illness and the loss of lives, while Covid-19 is marked by widespread containment measures and relatively lower fatalities among young people. The short-term costs of Covid-19 will probably dwarf those of past epidemics, due to the unprecedented and synchronised global sudden stop in economic activity induced by containment measures. The current estimated impact on global GDP growth for 2020 is around -4%, with substantial downside risks if containment policies are prolonged. Output losses are larger for major economies.

This paper studies the relationship between bank geographic complexity and risk. We use a unique dataset of 96 bank holding companies around the world to measure the geographic dispersion of their affiliates. We study how this dispersion interacts with economic and regulatory conditions to affect the riskiness of the bank.

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.

Our ongoing celebration of the Library of Congress National Book Festival continues with Poets Laureate Tracy K. Smith and Robert Hass discussing the making of poetry, the position of Poet Laureate and their new books, "American Journal: Fifty Poems for Our Time" (edited by Smith) and "A Little Book on Form: An Exploration into the Formal Imagination of Poetry" (Hass), on the Poetry & Prose stage at the 2018 Festival.

Our ongoing celebration of the Library of Congress National Book Festival continues with presidential scholar Annette Gordon-Reed discussing "The Hemingses of Monticello: An American Family" on the Special Programs stage at the 2015 Festival.

Our ongoing celebration of the Library of Congress National Book Festival continues with crime writer Patricia Cornwell discussing "Red Mist,” her 19th Kay Scarpetta novel, on the Fiction & Mystery stage at the 2012 Festival.

Our ongoing celebration of the Library of Congress National Book Festival continues with young adult and fantasy author Leigh Bardugo discussing "Crooked Kingdom" on the Genre Fiction stage at the 2018 Festival. This post includes prompts for writing and thinking that young readers, families and teachers can use to explore the author and the author’s work—recommended for ages 13+.

Our ongoing celebration of the Library of Congress National Book Festival continues with former Secretary of State Condoleezza Rice discussing "Democracy: Stories from the Long Road to Freedom" on the Main Stage at the 2017 Festival.

Our ongoing celebration of the Library of Congress National Book Festival continues with two-time Pulitzer Prize-winning novelist Colson Whitehead discussing "Zone One" on the Poetry & Prose stage at the 2012 Festival.

Our ongoing celebration of the Library of Congress National Book Festival continues with historian Michael Beschloss discussing “Presidents of War” on the Main Stage at the 2019 Festival.

Our ongoing celebration of the Library of Congress National Book Festival continues with crime and mystery writer Karin Slaughter discussing "Broken," part of her Will Trent series, on the Fiction & Mystery stage at the 2010 Festival.

Our ongoing celebration of the Library of Congress National Book Festival continues with actor and author Neil Patrick Harris discussing "The Magic Misfits: The Minor Third." The event, part of the new year-long National Book Festival Presents series, took place in the Library’s Coolidge Auditorium. This post includes prompts for writing and thinking that young readers, families and teachers can use to explore the author and the author’s work—recommended for ages 8+.

Even celebrity chef Julia Child said that the sleek appliance made mixing 'marvelous'

Rural educator Frank Cyr had the vision and pull to force the nation to standardize the color of the ubiquitous vehicle

A new book explores the evolution of cartography throughout more than a century of commercial air travel

Application deadline: 17 May 2020 | Banking Settlements Centre | Location: Basel, Switzerland

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.

Press release about the Basel Committee launching the consolidated Basel Framework, 16 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)

BIS Press Release - Shaping the future of payments: BIS Quarterly Review, 1 March 2020

Press release about Basel Committee assesses members' implementation of the Net Stable Funding Ratio and large exposures framework, 19 March 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 sets out additional measures to alleviate the impact of Covid-19", 3 April 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

CPMI Press release "Jurisdictions move towards full implementation of standards for financial market infrastructures", 8 April 2020.

BCBS Press release "Basel Committee issues progress report on banks' implementation of the 'Principles for effective risk data aggregation and reporting'", 29 April 2020

The American Geriatrics Society annually honors researchers, clinicians, educators, and emerging health professionals who have made outstanding contributions to high-quality, person-centered care for older people.

Acute myeloid leukemia is an aggressive cancer of the blood-forming system. It affects the hematopoietic stem cells, or blood stem cells, of various white blood cells and of the red blood cells and platelets.

With a discovery that could rewrite the immunology textbooks, an international group of scientists, including the teams of Bart Lambrecht, Martin Guilliams, Hamida Hammad, and Charlotte Scott (all from the VIB-UGent Center for Inflammation Research) identified a new type of antigen-presenting immune cell.

In a pandemic, it’s not just truth against misinformation. Every choice is an experiment.

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.

Holland America and Princess cruise lines issued notices this week cancelling the remainder of the season in Atlantic Canada, citing global health concerns, and that's taking a huge bite out of the Port of Sydney's budget.

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.

Police say efforts to find the driver of an abandoned car in Kings County last week likely saved his life.

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