Sentences with phrase «blocking buffer»

Membranes were blocked with LI - COR blocking buffer (LI - COR Biosciences, Lincoln, NE).

Membranes were blocked with Odyssey Blocking Buffer (LI - COR Biosciences) for 1 hour and incubated overnight at 4 °C with the respective Abs.

Sections were blocked for 1 h at RT in blocking buffer (PBS supplemented with 1 % w / v BSA, 0.3 % Triton X-100, and 1 % v / v donkey serum), which was used to dilute primary and secondary antibodies in later steps.

After three washing steps with TBS - T, a horseradish peroxidase antibody (diluted 1 ∶ 10,000 in blocking buffer) was used as secondary antibody.

The gels were transferred to Immobilon - FL PVDF membranes (EMD Millipore), and the membranes were blocked for 1 h in 50 % Odyssey blocking buffer (LI - COR Biotechnology).

Sections were blocked for 1 h at RT in blocking buffer [PBS supplemented with 1 % w / v bovine serum albumin (BSA), 0.3 % Triton X-100, and 1 % donkey serum], which was also used to dilute primary and secondary antibodies in subsequent steps.

To block unspecific staining, slides were incubated at room temperature for 15 min in ACE blocking buffer.

Samples were incubated with primary antibody (1/300 in blocking buffer) for 12 hours at 4 °C.

Isolated retinas were blocked and permeabilized with retina blocking buffer (10 % normal goat serum, Dako, #X0907, in 0.5 % Triton - X 100 and 1 % BSA in PBS) for 1 h at RT..

After washing with PBS and blocking for 30 minutes (IHC / ICC Blocking Buffer - Low Protein; eBioscience), cells were incubated with anti - Oct ‑ 4 antibody (diluted 1:150; eBiosicence) or an IgG2a K isotype control (diluted 1:150; eBiosicence) for one hour.

De Blasio, who previously said he would not march if Lopez Rivera was honored, nonetheless marched but kept a roughly 10 - block buffer between himself and the divisive Lopez Rivera.

Following the provided protocol, a portion of the RNA - containing lysate (5 — 10 µl) was hybridized at 54 degrees C overnight to RNA specific magnetic capture beads in the presence of blocking buffers, proteinase K and preordered mRNA probe sets specific for the genes of interest: HMOX1, Nrf2, Keap1, Bach1, and GAPDH.

Not exact matches

The rule maintains a long - established 100 - foot buffer zone that blocks coal mining near streams, but imposes stricter guidelines for exceptions to the 100 - foot rule.

A federal judge has ordered the creation of a «buffer zone» to keep protesters out of an area in front of the centre to prevent them from blocking access to the surgical centre.

Designed with a buffer between the filters, the mechanism automatically isolates the coffee grinds and tea leaves to block further extraction, ensuring consistent taste from the first sip to the last.

The Downers Grove Park District Board of Commissioners voted 4 - 1 last year to sue the hospital to block construction of a $ 14 million health and fitness facility, which environmentalists said would cause extensive ecological damage to Lyman Woods and a buffer zone.

Last month the Downers Grove Park District Board of Commissioners voted 4 - 1 to sue the hospital to block construction of the $ 14 million center, which environmentalists contend would cause extensive ecological damage to Lyman Woods and a buffer zone.

The membranes were blocked in 5 % nonfat milk (NFM) protein in Tris - buffered saline containing 0.05 % Tween (TBS - T).

After being washed with 4 × SSC (0.15 M NaCl in 0.015 M sodium citrate), the slides were treated with 20 μg / mL RNaseA at 37 °C for 30 min, washed with RNase buffer (0.5 M NaCl, 0.01 M Tris - HCl) three times, 0.5 × SSC twice, and 1 × buffer1 (0.1 M Tris and 0.15 M NaCl) once, and then incubated in Roche blocking solution for 30 min.

The tissue was paraformaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed before the tissue was blocked and incubated with the antibody for 1 hour.

Sections were incubated in 0.3 % hydrogen peroxide in PBS for 20 min, washed in PBS - T (0.01 M phosphate - buffered saline, 0.2 % Triton X-100) and blocked 1 h with 10 % normal goat serum (NGS) in PBS - T.

Tissue was fixed with paraformaldehyde and blocked with 5 % serum for 1 hour at 21 °C; antigen retrieval was by heat mediation in a citrate buffer.

Tissue was fixed with 10 % buffered formalin and blocked with 5 % serum for 60 minutes at 21 °C; antigen retrieval was by heat mediation in a 10mM sodium citrate buffer pH6.

IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4 % for 5 minutes, permeabilised with 0.5 % triton - PBS for 5 minutes, blocked with 5 % milk / 0.2 % tween for one hour.

Briefly, plates were coated overnight at 4 °C with the FMP2.1 recombinant AMA1 antigen (100 µL / well, 0.5 µg / mL), after which they were blocked with a 0.5 % boiled casein buffer for 1 hour at 22 °C.

The sections were treated with 3 % H2O2 in phosphate buffer saline (PBS), then incubated with serum blocking reagent for 30 minutes.

The plates were blocked with 2 % Marvel milk powder in phosphate buffered saline (PBS) for 2 h at 37 °C, washed three times with PBS, 0.05 % Tween 20.

Tissue samples were fixed by perfusion with paraformaldehyde, permeablized with 0.5 % Triton X-100 and blocked with 10 % serum for 1 hour at 25 °C; antigen retrieval was by heat mediation in 10 mM citrate buffer, pH 6, for 20 minutes at 97 °C in a water bath.

Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10 min at 70 °C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5 % BSA and probed with unpurified ab51608.

Tissue was fixed with formaldehyde and blocked with 10 % serum for 30 minutes at 22 °C; antigen retrieval was by heat mediation in a citrate buffer.

Sections were PFA fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH 6) prior to blocking in 10 % serum for 10 minutes at 25 °C.

Use a scrub brush to clean underneath the nails, and then gently buff them (our favorite buffer is Creative Nail Design Glossing Buffer Block, $ 5.50) to prepare the nails for pbuffer is Creative Nail Design Glossing Buffer Block, $ 5.50) to prepare the nails for pBuffer Block, $ 5.50) to prepare the nails for polish.

Items that cant be disinfected — nail - buffer blocks, nail brushes, emery boards — should be thrown away immediately after being used.

Since that first spark of inspiration became something tangible, significant others, buddies from work and siblings across the state act as sounding boards, therapists encouraging you back to the drawing board, and buffers through all those aggravating moments of writers block.

Instead of mounting the scratching pole directly to the hardwood floor, I created tension with a large screw at the top (using the face of the smaller file drawer front as a buffer between it and the ceiling) and secured the pole to a 2x4x4 block covered in carpet scraps.

Topics discussed in this batch include making decisions in the neutral, instant while running technique, effective sidestepping, block punishing attacks, crush moves, button buffering and frame advantage.

A Wild and Scenic designation creates a protected buffer along both sides of a river, blocks dams and other harmful water projects, and preserves a river's free - flowing nature.

Only a block away is the very busy Corrientes Avenue, and the huge trees inside the inner park of the complex acted as a buffer for street noise.

KyberNetwork is block - instantaneous and it significantly reduces the buffer between when an order is made and when it is filled.

Practitioners should receive training specifically about how: (a) to create a buffer zone to facilitate crossing the co-parental boundary, (b) to help the child separate from the enmeshment with the alienating parent, (c) to block intrusions from the alienating parent, (d) to strengthen the bond with the targeted parent, and (e) to weaken the coalition around the alienating parent — child dyad (Ellis and Boyan, 2010).

«Basically, you block your days into three kinds of blocks — strategic blocks, buffer blocks, and breakout blocks,» he explains.

What's more, less than 10 percent of total tenants will see their leased space expire each year through 2020, buffering Cousins Properties from seeing large blocks of empty space that weigh on net income.