Np14 bsa

Manufactured by Biosearch Technologies

NP14-BSA is a laboratory product manufactured by Biosearch Technologies. It serves as a reagent for various applications in scientific research. The core function of NP14-BSA is to provide a stable and standardized nanoparticle-protein conjugate solution.

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Lab products found in correlation

Multiscreen filter plate, by Merck Group (1 mentions) Bcip nbt plus substrate, by Mabtech (1 mentions) Microplate reader, by Tecan (1 mentions) Imject, by Thermo Fisher Scientific (1 mentions) Np27 cgg, by Biosearch Technologies (1 mentions) Np2 bsa, by Biosearch Technologies (1 mentions) Maxisorp elisa plate, by Thermo Fisher Scientific (1 mentions) A10551, by Thermo Fisher Scientific (1 mentions) Elispot plate, by Merck Group (1 mentions)

5 protocols using np14 bsa

NP-Specific Antibody ELISA and ELISPOT

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4-hydroxy-3-nitrophenylacytyl (NP)-specific IgM and IgG1 antibodies were determined by ELISA, and IgG1 PC formation was detected by ELISPOT, following previously published protocols.
For ELISA assay, 384-well flat-bottomed plates (NUNC) were coated at 4 °C overnight with 5 μg/mL NP₁₄-BSA (Biosearch Technologies), washed 3 times and blocked for 2 h at RT with 2% BSA in PBS (blocking buffer). Sera sample were serially diluted, added to each well, and subsequently incubated for 2 h at RT. After washing thoroughly for 5 times, biotinylated anti-IgM and anti-IgG1 antibodies (1: 1000 dilution in blocking buffer) were supplemented to each well for 1 h then conjugated with streptavidin-HRP. After adding in 3,3′,5,5′ tetramethyl benzidine thiobarbituric acid substrate, data were collected using tecan microplate reader (Tecan).
To carry out ELISPOT assay, NP₁₄-BSA (50 μg/ml, Biosearch Technologies) was used to coat the multiScreen filter plates (Millipore) at 4 °C overnight. Thereafter, 2% BSA in PBS was used to block the plate for 2 h at 37 °C. Single cells were added to each well and incubated for at least 2 h in a humidified incubator containing 5% CO₂. Subsequently, streptavidin–alkaline phosphatase (AKP)-conjugated anti-IgG1 antibodies were added to the filter plates for 1 h. BCIP/NBT-plus substrate (Mabtech) was used to develop the plate.

Li Y.F., Xu S., Huang Y., Ou X, & Lam K.P. (2017). Tyrosine kinase c-Abl regulates the survival of plasma cells. Scientific Reports, 7, 40133.

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Spag6 Knockout Immune Response

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C57/SV129 WT were irradiated and reconstituted i.v with 5 million bone marrow cells from WT or Spag6KO as previously described40 (link)41 (link). After 6 weeks, mice were footpad and i.p immunized with 10 μg 4-hydroxy-3-nitrophenylacetyl coupled to keyhole limpet hemocyanin at a ratio of 27:1 (Bioresearch Technologies) in 4 mg alum. Mice were bled at day 7 and organs harvested at day 14. NP-specific IgM and IgG1 was determined by ELISA using NP-14–BSA (15 μg/ml; Biosearch Technologies) as previously described42 (link).

Cooley L.F., El Shikh M.E., Li W., Keim R.C., Zhang Z., Strauss J.F., Zhang Z, & Conrad D.H. (2016). Impaired immunological synapse in sperm associated antigen 6 (SPAG6) deficient mice. Scientific Reports, 6, 25840.

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Alum-Precipitated NP-CGG Immunization

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For TD antibody responses, mixed BM chimeras were immunized i.p. with 50 µg NP₂₇-CGG in PBS (Biosearch Technologies) precipitated in alum (Imject; Thermo Fisher Scientific; 3:1 ratio). Alum mixed with PBS was used as a control. The production of NP-specific and total immunoglobulins (IgM and IgG1) was monitored by ELISA, using 96-well Maxisorp ELISA plates pretreated with 50 µl/well of 5 µg/ml NP_14.-BSA (Biosearch Technologies). Antibody isotypes and subclasses were detected with horseradish peroxidase–conjugated mouse IgM and subclass IgG-specific antibodies (Southern Biotech).

Jacque E., Schweighoffer E., Visekruna A., Papoutsopoulou S., Janzen J., Zillwood R., Tarlinton D.M., Tybulewicz V.L, & Ley S.C. (2014). IKK-induced NF-κB1 p105 proteolysis is critical for B cell antibody responses to T cell–dependent antigen. The Journal of Experimental Medicine, 211(10), 2085-2101.

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Hapten-specific ELISA for Antibody Profiling

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For hapten NP-specific enzyme-linked immunosorbent assay (ELISA), NP2-BSA (bovine serum albumin) or NP14-BSA (10 μg/ml; LGC Biosearch Technologies) was coated in PBS. NP2-BSA was used to detect high-affinity Abs, and NP14-BSA was used to detect high- and low-affinity Abs. After blocking with 3% FCS–RPMI 1640, diluted serum samples were added into the plates. Anti-IgG Abs (1:3000 dilution; Rockland), anti-IgG1 Abs (1:500 dilution; A10551; Invitrogen), anti-IgG2b Abs (1:500 dilution; SL257799, Invitrogen), and anti-IgG2c Abs (1:1000 dilution; PA1-29288, Invitrogen) were used for the detection of each isotype. For the virus HA and nuclear protein–specific ELISA, recombinant H3N2 influenza A virus HA protein or recombinant H3N2 influenza A virus nuclear protein (2 μg/ml) was coated on the MaxiSorp ELISA plate (Thermo Fisher Scientific) in KPL coating solution (Seracare/LGC Clinical Diagnostics, Inc.). After blocking with 0.5% milk powder in 0.1% Tween 20 containing PBS, diluted serum samples were added into the plates. Concentrations of each isotypes were analyzed by area under the curve (AUC).

Horiuchi S., Wu H., Liu W.C., Schmitt N., Provot J., Liu Y., Bentebibel S.E., Albrecht R.A., Schotsaert M., Forst C.V., Zhang B, & Ueno H. (2021). Tox2 is required for the maintenance of GC TFH cells and the generation of memory TFH cells. Science Advances, 7(41), eabj1249.

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Antigen-Specific ELISpot Assay for Splenic or Bone Marrow Cells

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For antigen-specific ELISpot with splenic or BM cells, 96-well ELISpot plates (Millipore) were coated overnight at 4 °C with NP3– or NP14–BSA (Biosearch Technologies), diluted in carbonate buffer. Plates were washed with PBS and blocked for 3 h at 37 °C with 10% FCS/B cell medium (BCM). 5 × 10⁵ cells were seeded per well and incubated for 24 h at 37 °C. Plates were washed with PBS-T (PBS with 0.025% Tween20). Biotinylated antibodies against the detecting isotype were diluted in 1% BSA/PBS and incubated for 2 h at 37 °C. After washing with PBS-T, streptavidin horseradish peroxidase Avidin D (Vector) in 1% BSA/PBS was added and incubated for 45 min at room temperature. For the development of spots, a developing buffer was prepared as follows: one tablet of each 3,3′- Diaminobenzidin peroxidase-substrate (0.7 mg/ml, Sigma-Aldrich, gold and silver tablets) was dissolved in 5 ml distilled water, the solution was pooled and filtered. 50 µl/well of this buffer was incubated for 8 min (splenocytes) or 12 min (BM cells). The reaction was stopped by washing with distilled water. Spots were visualized and counted with the ImmunoSpot Series 5 UV Analyzer (equipped with software version 5.0, CTL Europe).

Babushku T., Lechner M., Ehrenberg S., Rambold U., Schmidt-Supprian M., Yates A.J., Rane S., Zimber-Strobl U, & Strobl L.J. (2024). Notch2 controls developmental fate choices between germinal center and marginal zone B cells upon immunization. Nature Communications, 15, 1960.

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