Log In Sign up
The largest database of trusted experimental protocols

Para nitrophenyl phosphate substrate

Manufactured by Merck Group
Visit Supplier
Sourced in France, Panama, United States

Para-nitrophenyl phosphate substrate is a colorimetric substrate that can be used to detect and measure the activity of phosphatase enzymes. It undergoes a color change when cleaved by phosphatases, allowing for quantification of enzyme activity.

Automatically generated - may contain errors

Lab products found in correlation

Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Recombinant pa, by List Biological Laboratories (1 mentions) Alkaline phosphatase conjugated goat anti human igg, by Jackson ImmunoResearch (1 mentions) Emax plus microplate reader, by Molecular Devices (1 mentions) Microplate reader, by Agilent Technologies (1 mentions) Elisa plate, by Corning (1 mentions) Ovalbumin (ova), by Merck Group (1 mentions) Infinite 200 pro plate reader, by Tecan (1 mentions) Glacial acetic acid, by Thermo Fisher Scientific (1 mentions) Glycine buffer, by Merck Group (1 mentions) Penicillin streptomycin antibiotic solution, by Thermo Fisher Scientific (1 mentions) Propidium iodide, by Thermo Fisher Scientific (1 mentions) Alamar blue, by Thermo Fisher Scientific (1 mentions) Fluorescein diacetate fda, by Thermo Fisher Scientific (1 mentions) Oil red o, by Merck Group (1 mentions) Fitc albumin, by Merck Group (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Alizarin red s, by Merck Group (1 mentions) Ascorbic acid, by Merck Group (1 mentions)

8 protocols using para nitrophenyl phosphate substrate

1

Anti-collagen IgG ELISA in CIA Model

Check if the same lab product or an alternative is used in the 5 most similar protocols
Anti‐collagen IgG antibodies were evaluated by ELISA in B6 mice. After cCII adsorption on microtitration plates, mice sera, diluted at 1/1000 was incubated on coated plates. Anti‐cCII IgG antibodies were detected with alkaline phosphatase‐linked rat anti‐mouse IgG antibody (Sigma‐Aldrich, Saint‐Quentin Fallavier, France) after incubation with its para‐nitrophenylphosphate substrate (Sigma‐Aldrich) and read at 405 nm. Sera of CIA mice with severe disease from previous experiments was used as a positive control.
Denys A., Clavel G., Lemeiter D., Schischmanoff O., Boissier M, & Semerano L. (2016). Aortic VCAM‐1: an early marker of vascular inflammation in collagen‐induced arthritis. Journal of Cellular and Molecular Medicine, 20(5), 855-863.
+ Open protocol
+ Expand
2

Quantitative Antibody Assay for PA and HBs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Ninety-six-well microtiter plates (Corning, Lowell, MA) were coated with 1 μg/well of recombinant PA (List Biological Laboratories, Campbell, CA) or 0.02 μg/well recombinant HBs (Cell Sciences, Canton, MA). Diluted plasma was added, followed by anti-human IgG conjugate (Jackson ImmunoResearch, West Grove, PA) and para-nitrophenylphosphate substrate (Sigma-Aldrich, St. Louis, MO), with washing between steps. The optical density (OD) was detected using a Dynex MRX II microplate reader (Dynex Technologies, Chantilly, VA). Concentration of PA and HBs antibodies were calculated from standard curves of the reference sera AVR801 (BEI Resources, Manassas, VA) [27 (link)] and WHO international standard 07/164 (NIBSC, Potters Down, UK), respectively. Greater than 10 IU/L of anti-HBs is defined as positive or protected, and a good response is defined as 100 IU/L [8 (link), 28 (link)]. Non-human primate studies estimate the level of anti-PA IgG that predicts 80% survival in the year following a 3 dose priming series varies from 5.4–97.3 μg/mL depending on the time since last vaccination [29 (link)]. Therefore, individuals with anti-PA IgG < 5.4 μg/mL, 5.4–97.3 μg/mL, and >97.3 μg/mL are considered negative, equivocal, and positive, respectively.
Garman L., Vineyard A.J., Crowe S.R., Harley J.B., Spooner C.E., Collins L.C., Nelson M.R., Engler R.J, & James J.A. (2014). Humoral responses to independent vaccinations are correlated in healthy boosted adults. Vaccine, 32(43), 5624-5631.
+ Open protocol
+ Expand
3

EBNA411-426 Peptide Antibody Purification and ELISA

Check if the same lab product or an alternative is used in the 5 most similar protocols
EBNA411–426 peptide (EADYFEYHQEGGPDGE) was synthesized as a multiple antigenic peptide (MAP™) by the Oklahoma Molecular Biology Core Facility at the University of Oklahoma Health Science Center. The peptide was bound to cyanogen pre–activated Sepharose 4B [27 (link)]. To absorb antibodies, sera were recirculated (2–3 times) and concentrated to volume. Anti-EBNA411–426 antibodies were eluted with 3M sodium thiocyanate.
IgG responses to EBNA411–426 were measured by ELISA. Microliter plates were coated with 10µg/ml peptide in carbonate buffer overnight. Following addition of blocking buffer (0.1% BSA), serum samples diluted 1:100 in diluent (0.1% BSA, 0.05% Tween-20) were added and incubated for 2 hours at room temperature. Alkaline phosphatase conjugated goat anti-human IgG (Jackson Immunoresearch) was used as secondary antibody. The plates were developed using 1mg/ml para-nitrophenyl phosphate substrate (Sigma) in substrate buffer (0.1M glycine, pH 10.4, 1mM MgCL2, 1mM ZnCl2). The plates were read at 405nm on Emax Plus microplate reader (Molecular Devices).
Jog N.R., McClain M.T., Heinlen L.D., Gross T., Towner R., Guthridge J.M., Axtell R.C., Pardo G., Harley J.B, & James J.A. (2019). Epstein Barr Virus Nuclear Antigen 1 (EBNA-1) peptides recognized by adult multiple sclerosis patient sera induce neurologic symptoms in a murine model. Journal of autoimmunity, 106, 102332.
+ Open protocol
+ Expand
4

Capsule Polysaccharide Detection by ELISA

Check if the same lab product or an alternative is used in the 5 most similar protocols
To detect the capsule polysaccharide (PS) during purification, an inhibition-type enzyme-linked immunosorbent assay (ELISA) was performed. Briefly, the wells of ELISA plates (Corning Costar Corp., Acton, MA) were coated at 37°C with 1 μg/ml of 10A capsular PS (SSI, Copenhagen, Denmark) for 5 h in phosphate-buffered saline. After the plates were washed with PBS containing 0.05% Tween 20, 50-μl portions of the samples containing PS were added to the wells along with 50 μl of factor serum 10d (FS10d) at 1:10,000 dilutions. After 1 h of incubation at room temperature, the plates were washed and incubated for 1 h with 100 μl of alkaline phosphatase-conjugated goat anti-rabbit Ig (Southern Biotech) at 1:3,000 dilutions. The amount of the enzyme immobilized to the wells was then determined by incubating paranitrophenyl phosphate substrate (Sigma) in diethanolamine buffer for 1 to 2 h at room temperature and measuring the OD405 with a microplate reader (BioTek Instruments Inc., Winooski, VT). Purified 10A PS (SSI, Copenhagen, Denmark) was used as an assay standard (Fig. S5).
Ganaie F., Saad J.S., McGee L., van Tonder A.J., Bentley S.D., Lo S.W., Gladstone R.A., Turner P., Keenan J.D., Breiman R.F, & Nahm M.H. (2020). A New Pneumococcal Capsule Type, 10D, is the 100th Serotype and Has a Large cps Fragment from an Oral Streptococcus. mBio, 11(3), e00937-20.
+ Open protocol
+ Expand
5

ELISA for Detecting Anti-OVA Antibodies

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunolon II plates (ThermoScientific) were coated with 100 μg/ml ovalbumin (Sigma-Aldrich) and blocked with 1% bovine serum albumin and 0.05% NaN3. Sera were serially diluted from 1:50 dilution and assayed for anti-OVA reactivity against the plates by incubation for 2 hours at room temperature. Bound Ig was detected with a goat polyclonal anti-mouse Ig detection antibody conjugated to alkaline phosphatase (SouthernBiotech) and visualized at 405 nm using a para-nitrophenylphosphate substrate (Sigma-Aldrich) and Infinite 200 PRO plate reader (Tecan).
Nelsen M.K., Beard K.S., Plenter R.J., Kedl R.M., Clambey E.T, & Gill R.G. (2017). Disruption of Transplant Tolerance by an ‘Incognito’ Form of CD8+ T Cell-Dependent Memory. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 17(7), 1742-1753.
+ Open protocol
+ Expand
6

Synthetic Laponite Nanosilicates for Biomaterials

Check if the same lab product or an alternative is used in the 5 most similar protocols
Synthetic Laponite nanosilicates (NS), Laponite XLG (NS − F) and Laponite XL21 (NS + F), were kindly gifted by BYK, USA. Minimal essential medium –alpha modification (alpha MEM), fetal bovine serum (FBS), penicillin/streptomycin/antibiotic solution, Trypsin-EDTA, Phalloidin, fluorescein diacetate (FDA) and propidium iodide (PI), and alamar blue were purchased from Invitrogen, USA. BSA, β-glycerophosphate, ascorbic acid, dexamethasone, formaldehyde, Triton-X 100, FITC albumin, alizarin red S, Oil-O Red, and para-nitrophenyl phosphate substrate with glycine buffer were purchased from Sigma-Aldrich, USA. BCA Protein Assay Kit and Glacial acetic acid were purchased from Thermo Fisher Scientific, USA, and SRL, India. Anti-osteopontin, donkey antigoat IgG conjugated with Alexa Flour 488 purchased from Abcam, USA. DAPI, Alcian Blue was purchased from Alpha Aesar, USA. MTS assay kit was purchased from Promega, USA. Milli-Q water (18.2 MΩ.cm) was used wherever needed. All chemicals were used with no further modifications and purification.
Veernala I., Giri J., Pradhan A., Polley P., Singh R, & Yadava S.K. (2019). Effect of Fluoride Doping in Laponite Nanoplatelets on Osteogenic Differentiation of Human Dental Follicle Stem Cells (hDFSCs). Scientific Reports, 9, 915.
+ Open protocol
+ Expand
7

Macaque Serum IgG Anti-FHbp ELISA

Check if the same lab product or an alternative is used in the 5 most similar protocols
Serum IgG anti-FHbp titers were measured by ELISA (50 (link)). Briefly, recombinant FHbp ID 1 (2 μg/ml in phosphate-buffered saline) was added to the wells of a microtiter plate (Immulon 2 HB; Thermo Fisher), and the plate was incubated overnight at 4°C. After blocking, 5-fold serial dilutions of macaque serum starting at 1:500 were added and the plate was incubated for 1 h at room temperature. Bound macaque IgG was detected with goat anti-human IgG (Fc specific) conjugated to alkaline phosphatase (1:5,000; Sigma-Aldrich), which cross-reacts with macaque IgG. After 30 min of development with para-nitrophenyl phosphate substrate (1 mg/ml; Sigma), the optical density (OD) at 405 nm was measured in a plate reader.
Beernink P.T., Vianzon V., Lewis L.A., Moe G.R, & Granoff D.M. (2019). A Meningococcal Outer Membrane Vesicle Vaccine with Overexpressed Mutant FHbp Elicits Higher Protective Antibody Responses in Infant Rhesus Macaques than a Licensed Serogroup B Vaccine. mBio, 10(3), e01231-19.
+ Open protocol
+ Expand
8

Lectin Immunosorbent Assay for Glycan Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols
Lectin immunosorbent assay is a method akin to enzyme-linked immunosorbent assays, using lectins as probes to identify glycan structures.[24 (link)] Buffer composition was 10 mM phosphate-buffered saline (PBS) containing 150 mM sodium chloride, pH 7.2, at 4°C. The wells containing the synpatosomal lysates were washed several times with PBS containing 0.05% (w/v) Tween 20 and blocked with bovine serum albumin. A total of 100 μL of biotinylated Ulex europaeus agglutinin I lectin at 0.6 μg/mL (Vector Laboratories, Burlingame, CA) in PBS was added to each well and allowed to incubate for 1 hour. A total of 100 μL of 1:3000 dilution of 1 mg/mL streptavidin conjugated to alkaline phosphatase (Vector Laboratories) was added and signal was developed by incubation for 30 minutes with 100 μL of 1 mg/mL para-nitrophenyl phosphate substrate (Sigma-Aldrich). The reaction was stopped using 50 μL of 3 M sodium bicarbonate. Negative and positive controls were used for all of the experiments.
Cui H., Liu Y., Qin L., Wang L, & Huang Y. (2016). Increased membrane localization of pannexin1 in human corneal synaptosomes causes enhanced stimulated ATP release in chronic diabetes mellitus. Medicine, 95(49), e5084.
+ 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!