Log In Sign up
The largest database of trusted experimental protocols

Alkaline phosphatase conjugated to streptavidin

Manufactured by Vector Laboratories
Visit Supplier
Sourced in United States

Alkaline phosphatase conjugated to streptavidin is a commonly used detection reagent in various immunoassay and affinity-based techniques. It combines the specific binding properties of streptavidin with the enzymatic activity of alkaline phosphatase, allowing for the detection and amplification of target biomolecules.

Automatically generated - may contain errors

Lab products found in correlation

Sulfo nhs xbiotin, by Thermo Fisher Scientific (2 mentions) Sigmafast p nitrophenyl phosphate tablets, by Merck Group (2 mentions) Vector blue alkaline phosphatase substrate kit 3, by Vector Laboratories (1 mentions) Multiscreen 96 well filtration plate, by Merck Group (1 mentions) Np19 bsa, by Biosearch Technologies (1 mentions) Para nitrophenyl phosphate substrate, by Merck Group (1 mentions) Biotinylated secondary antibody, by Agilent Technologies (1 mentions) Methyl green, by Agilent Technologies (1 mentions) Amplex ultrared, by Thermo Fisher Scientific (1 mentions) Cyquant mtt cell viability assay, by Thermo Fisher Scientific (1 mentions) Nunc lab tek 8 well chamber slide, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)

6 protocols using alkaline phosphatase conjugated to streptavidin

1

Quantifying Antigen-Specific Antibody Responses

Check if the same lab product or an alternative is used in the 5 most similar protocols
Multi screen 96-well filtration plates (Millipore, Billerica, MA) were coated with 4-hydroxy-3-nitrophenyl acetyl conjugated to bovine serum albumin (NP19-BSA, Biosearch Technologies, Petaluma, CA) at a concentration of 10 μg/ml overnight at 4°C. Splenocyte suspensions from immunized mice were initially plated at 1 × 106 cells per well and then diluted serially (1:2) in RPMI1640 medium containing 10% FBS and incubated for six hours at 37°C. Bound antibodies were detected using biotinylated anti-mouse IgM and IgG Abs (Jackson Immunoresearch Laboratories, West, Grove, PA) followed by streptavidin conjugated to alkaline phosphatase (Vector Laboratories, Burlingame, CA). The plates were developed using the Vector Blue Alkaline–Phosphatase Substrate kit III (Vector Laboratories, Burlingame, CA). ELISpots were counted using a computerized imaging video system (Cellular Technology, Shaker Heights, OH).
Vuyyuru R., Shen S, & Manser T. (2015). Testing the role of the FcγRIIB immunoreceptor tyrosine-based inhibitory motif in regulation of the B cell immune response. Immunity, Inflammation and Disease, 3(3), 247-264.
+ Open protocol
+ Expand
2

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
3

Virus-like Particle ELISA Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Virus-like particles containing protease-purified Env trimers were prepared as previously described50 (link),51 (link),91 (link), and ELISAs on these VLPs were performed as described51 (link). Briefly, Immulon II plates were coated overnight at 4°C with VLPs at 20 times their concentration in transfection supernatants. Wells were washed with PBS and then blocked with 4% bovine serum albumin/10% fetal bovine serum in PBS. Various biotinylated monoclonal antibodies (biotinylated using sulfo-NHS-Xbiotin, Thermo), and CD4-IgG2 were then titrated in the presence or absence of a fixed concentration of 2μg/ml soluble CD4. Alkaline phosphatase conjugated to streptavidin (Vector Laboratories, Burlingame, CA; to detect biotinylated mAbs) or anti-Fc (Accurate, Westbury, NY; to detect CD4-IgG2) and SigmaFAST p-nitrophenyl phosphate tablets (Sigma) were then used to detect binding. Plates were read at 405 nm.
Kwon Y.D., Pancera M., Acharya P., Georgiev I.S., Crooks E.T., Gorman J., Joyce M.G., Guttman M., Ma X., Narpala S., Soto C., Terry D.S., Yang Y., Zhou T., Ahlsen G., Bailer R.T., Chambers M., Chuang G.Y., Doria-Rose N.A., Druz A., Hallen M.A., Harned A., Kirys T., Louder M.K., O’Dell S., Ofek G., Osawa K., Prabhakaran M., Sastry M., Stewart-Jones G.B., Stuckey J., Thomas P.V., Tittley T., Williams C., Zhang B., Zhao H., Zhou Z., Donald B.R., Lee L.K., Zolla-Pazner S., Baxa U., Schön A., Freire E., Shapiro L., Lee K.K., Arthos J., Munro J.B., Blanchard S.C., Mothes W., Binley J.M., McDermott A.B., Mascola J.R, & Kwong P.D. (2015). Crystal structure, conformational fixation, and entry-related interactions of mature ligand-free HIV-1 Env. Nature structural & molecular biology, 22(7), 522-531.
+ Open protocol
+ Expand
4

Annexin-A8 Expression in Wdr72 Mutant Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mandibles from Wdr72+/+, Wdr72+/−,Wdr72−/− litermates (n = 4) were immediately immerse-fixed in 4% paraformaldehyde (PFA)/0.06 M cacodylate buffer (pH 7.3) overnight and decalcified in 8% EDTA (pH 7.2) for 2 weeks at 4 °C. Samples were then dehydrated and paraffin-processed for routine embedding and sectioning. Sagittal incisor sections at 5 μm were deparaffinized and rehydrated, followed by incubation with 10% swine serum for blocking. Rabbit polyclonal antibody to annexin-A8 (Thermo Fisher Scientific) (PA5-31479) was used at 1:500 dilution. Following overnight incubation at 25 °C, sections were washed a biotinylated secondary antibody (Dako, Carpinteria, CA) was added for 1 h at 25 °C. Alkaline phosphatase conjugated to streptavidin (Vector Laboratories Inc., Burlingame, CA) was used to visualize the colorimetric reaction. Sections were then counterstained using methyl green (Dako, Carpinteria, CA).
Katsura K., Nakano Y., Zhang Y., Shemirani R., Li W, & Den Besten P. (2022). WDR72 regulates vesicle trafficking in ameloblasts. Scientific Reports, 12, 2820.
+ Open protocol
+ Expand
5

Virus-like Particle ELISA Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Virus-like particles containing protease-purified Env trimers were prepared as previously described50 (link),51 (link),91 (link), and ELISAs on these VLPs were performed as described51 (link). Briefly, Immulon II plates were coated overnight at 4°C with VLPs at 20 times their concentration in transfection supernatants. Wells were washed with PBS and then blocked with 4% bovine serum albumin/10% fetal bovine serum in PBS. Various biotinylated monoclonal antibodies (biotinylated using sulfo-NHS-Xbiotin, Thermo), and CD4-IgG2 were then titrated in the presence or absence of a fixed concentration of 2μg/ml soluble CD4. Alkaline phosphatase conjugated to streptavidin (Vector Laboratories, Burlingame, CA; to detect biotinylated mAbs) or anti-Fc (Accurate, Westbury, NY; to detect CD4-IgG2) and SigmaFAST p-nitrophenyl phosphate tablets (Sigma) were then used to detect binding. Plates were read at 405 nm.
Kwon Y.D., Pancera M., Acharya P., Georgiev I.S., Crooks E.T., Gorman J., Joyce M.G., Guttman M., Ma X., Narpala S., Soto C., Terry D.S., Yang Y., Zhou T., Ahlsen G., Bailer R.T., Chambers M., Chuang G.Y., Doria-Rose N.A., Druz A., Hallen M.A., Harned A., Kirys T., Louder M.K., O’Dell S., Ofek G., Osawa K., Prabhakaran M., Sastry M., Stewart-Jones G.B., Stuckey J., Thomas P.V., Tittley T., Williams C., Zhang B., Zhao H., Zhou Z., Donald B.R., Lee L.K., Zolla-Pazner S., Baxa U., Schön A., Freire E., Shapiro L., Lee K.K., Arthos J., Munro J.B., Blanchard S.C., Mothes W., Binley J.M., McDermott A.B., Mascola J.R, & Kwong P.D. (2015). Crystal structure, conformational fixation, and entry-related interactions of mature ligand-free HIV-1 Env. Nature structural & molecular biology, 22(7), 522-531.
+ Open protocol
+ Expand
6

Detailed Protocol for In-Vitro Cytotoxicity Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols
Dulbecco’s Modified Eagle Medium (DMEM), Amplex UltraRed, and 4-methylumbelliferyl phosphate (4-MUP) were obtained from Invitrogen (Carlsbad, CA, USA). The CYQUANT MTT Cell Viability Assay, which is based on the cellular conversion of 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) to an insoluble formazan product, Hoechst (H33342) dye, and Nunc Lab Tek 8-well chamber slides were from Thermo Fisher (Cambridge, MA USA). Alkaline Phosphatase Conjugated to Streptavidin was purchased from Vector Laboratories (Burlingame, CA, USA). Details pertaining to the Agent Orange chemicals are listed in Table 1, and antibodies used in immunoassays are listed in Table 2. All other fine chemicals were purchased from Thermo Fisher (Cambridge, MA USA), Sigma-Aldrich (St. Louis, MO, USA), or CalBiochem (Carlsbad, CA, USA).
de la Monte S.M., Goel A., Tong M, & Delikkaya B. (2023). Agent Orange Causes Metabolic Dysfunction and Molecular Pathology Reminiscent of Alzheimer’s Disease. Journal of Alzheimer's Disease Reports, 7(1), 751-766.
+ 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!