| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
The target of NP-BSA is the immune system itself, not a specific receptor. As a hapten-carrier conjugate, it functions as a complete antigen, eliciting an immune response against the NP hapten. B cells specific for the NP hapten bind to NP-BSA via their B cell receptors (BCRs), internalize the conjugate, and present NP-derived peptides on MHC class II molecules to CD4+ T helper cells. This T cell-dependent activation leads to antibody production, class switching, and affinity maturation. NP-BSA is used to study the mechanisms of B cell activation, T cell help, and antibody responses.
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| ln Vitro |
In vitro, NP-BSA is used in immunological assays to detect and characterize NP-specific B cells and antibodies. It is used in ELISA, ELISpot, and flow cytometry to measure NP-specific antibody titers and to identify NP-binding B cells. It is a tool to study B cell receptor (BCR) affinity maturation and T cell-dependent B cell activation. It is not cytotoxic at the concentrations used (1-10 ug/mL). It is a research reagent, not a therapeutic agent.
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| ln Vivo |
In vivo, NP-BSA is used as a model antigen to study T cell-dependent immune responses in mice. Immunization of mice with NP-BSA in adjuvant (e.g., alum, CFA) elicits a strong NP-specific antibody response, characterized by class switching, affinity maturation, and the generation of memory B cells. NP-BSA-immunized mice are used to study B cell development, plasma cell differentiation, and the mechanisms of humoral immunity. It is used in combination with NP-OVA (NP conjugated to ovalbumin) to study T cell-B cell interactions.
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| Enzyme Assay |
Not applicable. NP-BSA is a protein conjugate, not a standard enzyme inhibitor. Its purity and substitution ratio (number of NP groups per BSA molecule) are determined by UV spectrophotometry (A280 for protein, A430 for NP). The structure is confirmed by mass spectrometry. Physicochemical properties: MW ~66 kDa + NP, white to off-white powder, soluble in PBS and water. For ELISA, plates are coated with NP-BSA (1-10 ug/mL) overnight, blocked, and incubated with serum samples; bound antibodies are detected with anti-IgG-HRP.
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| Cell Assay |
For B cell staining, single-cell suspensions from spleen or lymph nodes of immunized mice are incubated with fluorescently labeled NP-BSA (e.g., NP-BSA-FITC, 1-10 ug/mL) for 30 min at 4degC. NP-specific B cells are identified by flow cytometry as B220+ NP+. For ELISpot, plates are coated with NP-BSA (10 ug/mL), and splenocytes are added; NP-specific antibody-secreting cells (ASCs) are detected. For proliferation assays, B cells are labeled with CFSE and cultured with NP-BSA and T cells; proliferation is measured by flow cytometry.
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| Animal Protocol |
For in vivo immunization, female BALB/c mice or C57BL/6 mice (6-8 weeks old, n=5-10/group) are immunized intraperitoneally (IP) or subcutaneously (SC) with 50-100 ug of NP-BSA emulsified in complete Freund‘s adjuvant (CFA, 1:1) on day 0. Booster injections (50 ug in incomplete Freund‘s adjuvant, IFA) are given on day 14 and/or day 28. Blood is collected on days 7, 14, 21, 28, 35, and 42. NP-specific serum antibody titers (IgM, IgG1, IgG2a, etc.) are measured by ELISA. For affinity maturation studies, serum is collected at multiple time points, and the relative affinity of NP-specific antibodies is measured by dissociation rate in the presence of the hapten NP-cap. Germinal center B cells are analyzed by flow cytometry of splenocytes (B220+ GL7+ Fas+).
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| ADME/Pharmacokinetics |
NP-BSA is not intended for pharmacokinetic studies as it is an antigen, not a drug. When injected IP or SC, the protein conjugate is taken up by antigen-presenting cells (APCs) in the draining lymph node and spleen. It is processed and presented to T cells. The protein is degraded over days. For research use, it is stored as a lyophilized powder at -20degC (powder: -20degC for 3 years, 4degC for 2 years; in solution: -80degC for 1 year), protected from light and moisture.
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| Toxicity/Toxicokinetics |
NP-BSA has low toxicity. Immunization of mice with NP-BSA in adjuvant is generally well-tolerated. However, the adjuvant may cause local inflammation. Standard safety precautions for handling biological reagents apply: use PPE (gloves, lab coat, safety goggles), work in a fume hood, avoid inhalation and skin contact. For research use only.
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| References | |
| Additional Infomation |
NP-BSA (4-Hydroxy-3-Nitrophenyl Acetate-Bovine Serum Albumin; CAS# not assigned) is a research-grade hapten-carrier conjugate used as a model antigen in immunological studies. It is not an FDA-approved drug. It is used for studying T cell-dependent B cell activation, affinity maturation, germinal center reactions, and vaccine development. For research use only, not for diagnostic or therapeutic applications. Storage: powder at -20degC for 3 years, 4degC for 2 years; in solvent at -80degC for 1 year.
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| Appearance |
Light yellow to yellow liquid
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| Synonyms |
4-Hydroxy-3-nitrophenylacetyl-bovine serum albumin
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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