| Field | Specification |
|---|---|
| Mfr No | |
| Assay Time | |
| Assay Type | |
| Detection Range | |
| Expression System | |
| Gene ID | |
| Immunogen | Expression system for standard: E.coli; Immunogen sequence: L80-L235 |
| Product Type | |
| Reactivity | |
| Sample Type(s) | cell culture supernatants, serum and plasma (heparin, EDTA) |
| Sensitivity | |
| Storage | |
| Target | |
| UniProt # |
Background
Mouse TNF Alpha/Tumor necrosis factor (Tnf) is an established target in many assay panels, supporting hypothesis testing across diverse biological systems. This target is frequently investigated in Molecular & Cellular Biology research contexts. Cytokines and chemokines act as soluble messengers that coordinate immune cell activation, trafficking, and effector functions. Their concentrations can change rapidly in response to infection, tissue injury, or immune stimulation.
Biological function and signaling context
In immune signaling networks, cytokine production is often induced by pattern-recognition pathways and inflammatory transcriptional programs, while feedback regulators can dampen responses to restore homeostasis. Chemokine gradients guide leukocyte migration, influencing which cell populations accumulate at a site and how long they persist.
Why it matters in research
- Immune activation readout: Shifts in abundance can reflect pathway engagement and cellular activation state.
- Microenvironment profiling: Levels can help characterize inflammatory tone in tissues or biofluids.
- Response monitoring: Time-course measurements support interpretation of stimulus, treatment, or infection models.
Disease and translational relevance
Many cytokines and chemokines are reported to associate with inflammatory, autoimmune, infectious, and oncology-related processes. In research settings, interpreting changes benefits from pairing this analyte with complementary markers (e.g., upstream triggers, downstream effectors, and cell-type indicators) and considering matrix effects.
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Sample data
| Concentration (pg/ml) | 0 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.051 | 0.090 | 0.156 | 0.281 | 0.443 | 0.723 | 1.284 | 1.958 |
Kit components
| Catalog number | Description | Quantity | Storage of opened/reconstituted material |
|---|---|---|---|
| EZ0527-CA | Mouse anti- mouse Tnf monoclonal antibody (Capture Antibody) | 500 µl, 4 mg/mL (recommended dilution 1:100) | Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. |
| EZ0527-DA | Biotinylated goat anti- mouse Tnf polyclonal antibody (Detection Antibody) | 500 µl, 0.15 µg/mL (recommended dilution 1:100) | |
| AR1103 | Avidin-Biotin-Peroxidase Complex (ABC) | 500 µl (recommended dilution 1:100) | |
| EK0527-ST | Lyophilized recombinant mouse Tnf standard | 10 ng/tube×3 | Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. |
Materials required but not provided
- Microplate reader in standard size.
- Automated plate washer.
- Incubator.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- 96 well microplate (Cat# AR1100)
- Plate Sealers.
- Capture Antibody Diluent: PBS.
- Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered.
- Substrate Solution: Tetramethylbenzidine (Cat# AR1104)
- Stop Solution: 2 N H2SO4 (Cat# AR1105)
- Wash Buffer (PBS and PBS-T). Recipes:
- PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 µm filtered.
- PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4.
- *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)
►How many plates does one EZ-Set™ kit cover?
Each EZ-Set™ kit contains antibody pairs and recombinant protein standards sufficient for 5 × 96-well plates (480 wells total), making it significantly more cost-effective per well than pre-coated kits for high-throughput users.
►What additional materials do I need to run an EZ-Set™ assay?
You will need: (1) a 96-well ELISA plate (e.g. Boster Cat# AR1100); (2) substrate solution (TMB, Cat# AR1104); (3) stop solution (2N H₂SO₄, Cat# AR1105); (4) wash buffer (PBS and PBS-T); (5) coating diluent (PBS); and (6) reagent diluent (1% BSA in PBS, pH 7.2–7.4). These are all included in the EZ Set Accessory Kit (Cat# EZA001), which can be purchased separately.
►How does EZ-Set™ differ from pre-coated PicoKine® kits?
EZ-Set™ kits provide the matched antibody pairs, HRP-conjugated Avidin-Biotin Complex (ABC), and recombinant protein standard — but not the pre-coated plate or accessory reagents. This gives experienced ELISA users flexibility to optimise coating conditions, plate choice, and buffer formulations. PicoKine® kits are fully configured for immediate use.
►What sensitivity can I expect from EZ-Set™ kits?
EZ-Set™ kits are designed to match PicoKine® performance. Sensitivity typically ranges from <2 pg/mL to <60 pg/mL depending on the target. For example, Human BMP-2 EZ-Set™ achieves <2 pg/mL and Human M-CSF EZ-Set™ achieves <10 pg/mL. Actual sensitivity in your lab depends on plate quality, coating efficiency, and protocol adherence.
►Can I use my own ELISA plate brand?
Yes — EZ-Set™ components can be used with ELISA plates from other manufacturers, provided they have adequate protein-binding capacity (high-binding plates recommended). Boster recommends their own Cat# AR1100 plates as they have been validated with EZ-Set™ antibody pairs.
►How do I optimise coating concentration and incubation conditions?
Start with the coating concentration recommended in the datasheet. If signal is weak, increase coating antibody concentration or extend coating time (overnight at 4°C rather than 2 hours at room temperature). If background is high, increase blocking time or BSA concentration. A checkerboard titration of capture vs. detection antibody concentrations is recommended when working with a new target.
►Are the kit components sterile?
Components are bottled under aseptic conditions but are not certified sterile. Filter through a 0.2 µm membrane if sterile reagents are required for your application.
►How do I analyse EZ-Set™ ELISA data?
Generate a standard curve by plotting OD 450 nm against the known standard concentrations and apply a 4-parameter logistic (4PL) curve fit. Back-calculate sample concentrations from the curve. Use Boster's free online ELISA data analysis tool at bosterbio.com/biology-research-tools/elisa-data-analysis-online or any curve-fitting software that supports 4PL regression.
Can’t Find What You’re Looking For? We can help you source the best match or customize an ELISA solution for your study. Options may include alternative target synonyms, different species reactivity, sample type/matrix compatibility (serum/plasma/lysate/supernatant), assay format (sandwich/competitive), sensitivity/range, detection chemistry (colorimetric/fluorescent/chemiluminescent), plate format (pre-coated/uncoated, strips vs full plate), and bulk or custom packaging. Click Talk to a Scientist to submit a request form, email us at support@biohippo.com, or explore our Research Services for additional support. Our team will be in contact with you shortly.
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