The largest database of trusted experimental protocols

High bind microplate

Manufactured by Corning
Sourced in United States

The High Bind Microplate is a laboratory equipment product designed for high-efficiency binding of proteins, peptides, and other biomolecules. It provides a high-affinity surface that promotes effective adsorption of target analytes. The microplate is suitable for a variety of immunoassay and biochemical applications that require efficient capture and detection of biomolecules.

Automatically generated - may contain errors

8 protocols using high bind microplate

1

Antibody Binding Assay on Microplate

Check if the same lab product or an alternative is used in the 5 most similar protocols
Wells of a 96 well white flat bottom polystyrene High Bind microplate (Costar) were coated overnight at 4°C with recombinant target protein (0.2 μg/mL in PBS) followed by a saturation step of at least 2 h at 37°C with gelatine 0.5% in PBS. NanoLuc fused antibody 1 nmol/L was then added in each well simultaneously with various amounts of competitors or controls. After 1 h incubation and three washing steps, 50 μL of Nano-GloTM assay substrate (1/400 dilution in PBS/BSA 0.1%) was added and luminescence read (0.1 s/well) on a Berthold Mithras LB940 microplate reader.
+ Open protocol
+ Expand
2

Quantifying RSV Binding to LL-37

Check if the same lab product or an alternative is used in the 5 most similar protocols
High affinity binding plates (High bind microplate, Costar Corning, Incorporate, Tewksbury, USA, #3590 were coated with 1×106 PFU RSV in coating buffer (15 mM Na2CO3, 35 mM NaHCO3, pH 9.6), control cell culture medium or 2.5% BSA for 12 h at 4°C, washed with PBS and incubated with 2.5% BSA for 3 h. Wells were exposed to 2 μg/mL LL-37 for 30 minutes at 4°C, washed with 0.02% Tween20/PBS. Bound LL-37 was detected by anti-LL-37 antibody (1:6400, #G-075-06, Phoenix pharmaceuticals, Burlingame, USA), HRP-labelled secondary antibody (1:5000, goat-anti-rabbit IgG (H+L) Jackson laboratories, Westgrove, USA, #111-035-045) and TMB substrate. Reaction was stopped using 0.5 M sulphuric acid and absorbance at A450 was determined using a plate reader (Synergy HT, Biotek, Winooski, USA).
+ Open protocol
+ Expand
3

Quantifying RSV-LL-37 Binding Interactions

Check if the same lab product or an alternative is used in the 5 most similar protocols
High-affinity binding plates [High bind microplate (Costar Corning, Tewksbury, MA) 3590 were coated with 1 × 106 PFU RSV in coating buffer (15 mM Na2CO3, 35 mM NaHCO3, pH 9.6)], control cell culture medium, or 2.5% BSA for 12 h at 4°C, washed with PBS, and incubated with 2.5% BSA for 3 h. Wells were exposed to 2 μg/ml LL-37 for 30 min at 4°C, washed with 0.02% Tween 20/PBS. Bound LL-37 was detected by anti–LL-37 Ab (1:6400, G-075-06; Phoenix Pharmaceuticals, Burlingame, CA), HRP-labeled secondary Ab [1:5000, goat anti-rabbit IgG (H+L) (Jackson Laboratories, West Grove, PA), 111-035-045], and TMB substrate. Reaction was stopped using 0.5 M sulfuric acid, and absorbance at A450 was determined using a plate reader (Synergy HT; Biotek, Winooski, VT).
+ Open protocol
+ Expand
4

ELISA Detection of TTSuV1 Antibodies

Check if the same lab product or an alternative is used in the 5 most similar protocols
Plates (High Bind Microplate, Corning®, Corning, NY) were coated with 50 μl of a 1:100,000 dilution of the purified TTSuV1 ORF2 the antigen. Coated plates were blocked with 2% BSA and 2% normal sheep serum in a commercial block (General block, ImmunoChemistry Technologies, Bloomington, MN) for 2 hrs at 37 °C, incubated with 1:50 of the human or swine sera for 2 hours at 37 °C, in duplicate, followed by the respective anti-species HRPO-conjugate (KPL, Gaithersburg, MD), at a 1:5000 dilution for 45 mins at 37 °C, and incubation with the substrate (TMB, KPL, Gaithersburg, MD). The reaction was stopped with a 1 M HCl solution after 2 mins. Plates were read at 450 nm on an ELISA plate reader (Elx800 reader, BioTek Instruments, Inc., Winooski, VT).
+ Open protocol
+ Expand
5

Quantifying Chimeric Antibody Titers

Check if the same lab product or an alternative is used in the 5 most similar protocols
The chimeric EG2-hFc monoclonal antibody (mAb) titers were determined by “sandwich” enzyme-linked immunosorbent assay (ELISA). A 96-well high-bind microplate (Corning, ref. 3369) was incubated overnight with goat anti-human Fc-specific antibodies (Sigma, ref. I2136) diluted 1:30,000 in PBS. Wells were washed three times with PBS containing 0.05% Tween 20 (Sigma, ref. P1379) and blocked for 1 h with a solution of 1% bovine serum albumin (BSA, Sigma, ref. A7030) in PBS. Extracellular medium samples were diluted 1:500 to 1:10,000 in the PBS-BSA solution, depending on the day of culture. The plate was washed and loaded with the diluted samples or human IgG whole molecule standards (Cederlane, ref. 009-000-003). After 1 h, the plate was washed and incubated for 1 h with peroxidase-conjugated goat anti-human Fc-specific antibodies (Sigma, ref. A0170). After washing, the plate was revealed with TMB substrate (Sigma, ref. T0440) and incubated for 20 min in the dark. Absorbance was read at 630 nm four times in the span of ten minutes and the slopes of optical density variations were compared to the standards by linear regression to determine the concentration in each well.
+ Open protocol
+ Expand
6

Mouse Mer, Axl, and Protein S ELISA

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mer and Axl were measured using Mouse Mer and Mouse Axl DuoSet ELISA (R&D Systems). To measure Protein S in plasma, polystyrene High Bind Microplates (Corning) were coated with a polyclonal anti-human Pros1 antibody (Dako) in 75 mM Na-carbonate pH 9.6 o/n at +4 °C. In between each assay step, plates were washed 3× with PBS containing 0.1% Tween 20. Wells were blocked with 2 % BSA in PBS, which was also used as sample and antibody dilution buffer. A standard curve was generated with a pooled plasma sample from 11 mice, and individual samples diluted at 1:400 were analyzed. Samples and standards were incubated on the plate for 2 h at RT, followed by incubation with a rat anti-mouse Pros1 antibody (R&D Systems) for 1 h. After an additional 1 h incubation with a goat anti-rat HRP conjugate (Jackson ImmunoResearch), the plates were developed with a TMB substrate (R&D Systems).
+ Open protocol
+ Expand
7

VEGF Binding Assay Using Heparan Sulfate

Check if the same lab product or an alternative is used in the 5 most similar protocols
Wells of 96-well High Bind microplates (Corning) were coated with fractionated heparan sulfate polymer (5 μg) or with VEGF capture Ab (100 ng) at RT for 16 h. Wells were then washed three times with wash buffer (R&D Systems), incubated with recombinant VEGF189 (2 ng) for 2 h, and washed again. Thereafter, bevacizumab or VEGFR1/R2-Fc (100 ng in a reaction volume of 100 μL) were added to wells that contained VEGF189 bound to heparan sulfate polymer or to VEGF capture Ab, and incubated for 2 h at RT. Wells were then washed three times. Binding of bevacizumab and VEGFR1/R2-Fc was detected by incubation with HRP-conjugated goat anti-human IgG (Fc-specific) (1:5000 dilution) for 1 h at RT, followed by washing and addition of HRP substrate solution (R&D Systems). Absorbance was read at 450 nm. The reading of wells that contained heparin-bound VEGF189 was normalized to the reading of wells that contained VEGF bound to VEGF capture Ab.
+ Open protocol
+ Expand
8

Screening Nanobody Clones from Phage Display

Check if the same lab product or an alternative is used in the 5 most similar protocols
Blocked recombinant phage supernatants (100 μl) were added to 96-well clear flat-bottom polystyrene High Bind microplates (Corning, USA) coated with 3 μg/ml Ptedn (to screen nanobody clones) or with 3 μg/ml BSA (negative control) for 1 h at 37 ℃. Microplates were rinsed 10 times with PBS/0.1% Tween 20 (PBST), incubated with 100 μl/ well of horseradish peroxidase (HRP)-conjugated anti-M13 antibody (1:5000; GE Healthcare, USA) for 1 h at 37 ℃, rinsed 10 times with PBST, and 100 μl/well of TMB singlecomponent substrate solution (Solarbio Science & Technology) was added in the dark for 10 min at room temperature. After the reaction was terminated with equal volume of 1 M H 2 SO 4 , optical density (OD) was measured at 450 nm and the positive clones with the highest OD values were selected for sequencing.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!