A planned exhibition at the Portland Art Museum highlights the boldness of their work

This little prince is eagerly awaiting the arrival of his Birthday guests. Unbeknownst to him, so is his feline friend.

This praying mantis looks completely shocked and like it was caught doing something wrong. I had to laugh while taking this shot, as mantis seem to be very expressive sometimes.

This photo shows the beautiful landscape of Capital Reef National Park with its amazing old orchards in foreground. Capital Reef has many old orchards of peaches, apples and apricots. The park lets you eat all you want if you eat it in the orchard.

The proboscis monkey is a species endemic to Borneo. I areas where they occur they are quite easy to encounter as they have reddish fur and make strange burping sounds for communication.

Spring bloom near my house

The National Trust for Historic Preservation is looking for 1,000 places tied to women's history, and to share the stories of the figures behind them

The question of bulletproofing vexed physicians and public figures for years, before pioneering inventors experimented with silk

While COVID-19 has us homebound, it’s a good time to reflect on the peculiar histories of housewares we take for granted

A new book explores how George Washington shaped the group of advisors as an institution to meet his own needs

The Kent State shootings further widened the chasm among a citizenry divided over the Vietnam War

The smart tool gives users step-by-step instructions based on WHO’s handwashing protocol

As spring begins, the surviving bees in the hive pick up their first task of the new year: dumping the corpses of the bees that died over winter.

The government of the Northwest Territories is releasing $8.7 million in federal funding to five airlines in the N.W.T. offering schedule-based passenger service.

The government of the Northwest Territories has made an exemption on border restrictions for a veterinarian from Alberta to practice in the territory, after he received public and political support.

Schools were closed March 13 to reduce the risk of spread of the coronavirus, and there is no plan to reopen them by the end of the current school year in June.

Many retailers are eager to hear more details from the province about when and how they should reopen after weeks of being closed to the public. The next phase of recovery is expected to start Friday.

The village of La Loche is calling on the provincial government to force the closure of liquor stores in the community, which is in the throes of a COVID-19 outbreak. 

Closing arguments were presented at the trial of Phillip Lionel Levac on Friday at Regina Court of Queen's Bench.

Changes are coming to Thunder Bay's popular Ribfest event because of the COVID-19 outbreak, organizers said.

It's been more than a week since there were any new or active cases of COVID-19 in the catchment area for the Northwestern Health Unit (NWHU), and staff can not focus increasing testing.

<span class="topsy_trackback_comment"><span class="topsy_twitter_username"><span class="topsy_trackback_content">Bebo doesn't fancy Liz Taylor! | RSS Feeds – IMC OnAir, India ...: May not do Madhur Bhandarkar's film which appar... http://bit.ly/9btJW3</span></span>

Job of The Week: CEO – Nordoff Robbins Music Therapy http://bit.ly/aFIMNM

<span class="topsy_trackback_comment"><span class="topsy_twitter_username"><span class="topsy_trackback_content">12 Cool India Facts [For President Obama's Visit] – RantRave ...: Washington Post12 Cool India Facts [For Presid... http://bit.ly/dmozLC</span></span>

<span class="topsy_trackback_comment"><span class="topsy_twitter_username"><span class="topsy_trackback_content">12 Cool India Facts [For President Obama's Visit] – RantRave ...: Washington Post12 Cool India Facts [For Presid... http://bit.ly/dmozLC</span></span>

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<span class="topsy_trackback_comment"><span class="topsy_twitter_username"><span class="topsy_trackback_content">Fresh trouble for SRK brought by the Shiv Sena men http://bit.ly/i8tocu #IMCRadio.net</span></span>

Newfoundland and Labrador's already fragile economy suffered another gut-punch in April, with Statistics Canada reporting Friday that 29,000 jobs were lost in April alone.

Green Depots will begin to reopen across the province on Monday, according to the Multi-Materials Stewardship Board, though dates and hours of operation may vary between sites. 

Demand for service is not expected to recover in the coming weeks, says Marine Atlantic.

La tripulación de técnicos del Logos Hope sabe que el mantenimiento del barco es una forma de hacer ministerio.

Durante un viaje misionero de un mes, Eduardo aprendió a escuchar y reconocer la voz de Dios, tanto personalmente como ministrando a otros.

The vessel, formerly known as the Edmonton Queen, got skewered by its own moorings last month as the icy North Saskatchewan River began to thaw.

Disaster relief funding for punishing spring floods is welcome news to civic leaders in northern Alberta.

A longstanding battle between an Alberta entrepreneur and a $50-million lottery winner is still raging after a new legal judgment, a securities investigation, allegations of harassment and even duelling ebooks.

In a new twist on oral history, University of Alberta paleontologists have discovered that an Argentinian reptile from 95 million years ago developed a type of tooth enamel that is common in humans and other mammals but rare among reptiles.

Every year Washingtonians are treated to a feast for the eyes as ornamental cherry trees bloom across the city, most prominently by the Tidal Basin. As cherry blossom season approaches, we would like to share information about two related resources that we hope will offer some inspiration for those near and far: a selected set […]

The following is a guest post by Katherine Blood, Curator of Fine Prints, Prints & Photographs Division. As the Library of Congress marks its 220th year of serving the nation, the publication of a new guide tells two stories: how staff have for decades worked with art professionals to build the collections and how by […]

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.

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.

Over many years, the Xenopus laevis embryo has provided a powerful model system to investigate how mechanical forces regulate cellular function. Here, we describe a system to apply reproducible tensile and compressive force to X. laevis animal cap tissue explants and to simultaneously assess cellular behavior using live confocal imaging.

Purified RNA may need to be concentrated by precipitation for downstream applications. Precipitation of RNA with ethanol (or isopropanol) is the standard method to recover RNA from aqueous solutions.

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.

Before probing blots for the presence of an antigen, the total composition of the transferred proteins can be determined by staining the nitrocellulose or polyvinylidene fluoride (PVDF) membrane. Staining for proteins is useful to determine the position of the non-prestained molecular weight markers or individual lanes on the gel and to ensure that efficient transfer has occurred. It can be also used to verify equal loading of the samples in the gel when a comparison of the protein of interest between the different samples is important. The conventional procedures such as Coomassie Blue and silver staining methods used for staining polyacrylamide gels are incompatible with immunoblotting. Ponceau S is the more common staining method in immunoblotting protocols because it is compatible with antibody–antigen binding, is cost efficient, and provides a good contrast between the stained bands and background. In this protocol, nitrocellulose or PVDF membrane is rinsed with ultrapure H₂O after the transfer of proteins. Ponceau S dye is applied as an acidic aqueous solution, and the proteins on the membrane are stained with red color. The membrane is briefly destained with water and can be photographed or scanned to obtain the image of the total protein staining. Individual lane positions or the molecular weight standards can be marked with a pencil, if required.

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.

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.

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.