Invitrogen™ Mouse IL-17E/IL-25 Uncoated ELISA Kit with Plates

Catalog No.	Quantity
50-171-75	2 x 96 Tests

Catalog No. 50-171-75 Supplier Invitrogen™ Supplier No. 88700222

Description

ELISA

The Mouse Interleukin 17E/Interleukin 25 (IL-17E) Uncoated ELISA Kit contains pre-matched antibody pairs, plates and reagents for performing quantitative enzyme linked immunosorbent assays (ELISA) to detect and quantify protein levels of mouse IL-17E. Wash Buffer and Stop Solution are needed to complete the ELISA reaction and are sold separately. Principle of the method ELISAs are designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody is coated to the bottom of the wells of a microplate, which is an overnight process. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. A sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.

Interleukin 25 (IL-25) is a member of the IL-17 family and is identified as a proinflammatory cytokine which is produced by Th17 effector cells thus enabling the Th17 subset to initiate or maintain the autoimmune process in diseases such as Rheumatoid Arthritis. IL-25 also induces activation of NF-kappa-B and stimulates production of the proinflammatory chemokine IL-8. IL-25 itself is activated through IL-23, IL-5, and TGF-beta.

Specifications

• Accession Number: Q8VHH8
• Assay Range: 32-2000 pg/mL
• Assay Sensitivity: 32 pg/mL
• Content And Storage: Capture Antibody: Pre-titrated, purified antibody
  Detection Antibody: Pre-titrated, biotin-conjugated antibody
  Standard: Recombinant cytokine for generating standard curve and calibrating samples
  Coating Buffer: 10X PBS ELISA Coating Buffer
  Assay Diluent: 5X Concentrated
  Sample Diluent B: 12 mL of a 1X solution per plate
  Detection Enzyme: Pre-titrated Avidin-HRP
  Substrate Solution: Tetramethylbenzidine (TMB) Solution
  Certificate of Analysis: Lot-specific instructions for dilution of antibodies and standards.
  96-well Plates: Corning Costar flat-bottom plates (included with Cat. Nos. ending in -22 and -86)
• Detectable Analytes: IL-17 E (IL-25)
• Assay: 16-2,000 pg/ml
• Conjugate: HRP
• Immunoassay Kit Format: Uncoated ELISA Kit with Plates
• Product Type: ELISA
• Sample Type: Serum, Supernatant
• Target Species: Mouse
• For Use With (Application): ELISA
• For Use With (Equipment): Absorbance Microplate Reader
• Gene Alias: D17Wsu104e; il 17e; Il17e; IL-17E; Il25; IL-25; ILN; Interleukin; interleukin 17E; interleukin 25; interleukin-17E; Interleukin-25; Ly6elg; Mydgf; myeloid derived growth factor; myeloid-derived growth factor; RGD1561632; UNQ3120/PRO10272; UPF0556 protein C19orf10 homologD17Wsu104e; il 17e; Il17e; IL-17E; Il25; IL-25; ILN; Interleukin; interleukin 17E; interleukin 25; interleukin-17E; Interleukin-25; Ly6elg; Mydgf; myeloid derived growth factor; myeloid-derived growth factor; RGD1561632; UNQ3120/PRO10272; UPF0556 protein C19orf10 homolog; BSF-2; BSF2; CDF; HGF; HSF; IFN-beta-2; IFNB2; IL-6
• Gene ID (Entrez): 140806
• Gene Symbol: IL25
• Host Species: Mouse
• Kit Contents: Capture Antibody: Pre-titrated, purified antibody, Detection Antibody: Pre-titrated, biotin-conjugated antibody, Standard: Recombinant cytokine for generating standard curve and calibrating samples, Coating Buffer: 10X PBS ELISA Coating Buffer, Assay Diluent: 5X Concentrated, Sample Diluent B: 12 mL of a 1X solution per plate, Detection Enzyme: Pre-titrated Streptavidin-HRP, Substrate Solution: Tetramethylbenzidine (TMB) Solution, Certificate of Analysis: Lot-specific instructions for dilution of antibodies and standards 96 Well Plate: High affinity 96-well plate (included with product Cat. Nos. ending in suffixes -22 and -86), Purified antibody, Detection Antibody: Pre-titrated, Biotin-conjugated antibody, Standard: Recombinant cytokine for generating standard curve and calibrating samples, Coating Buffer: 10X PBS ELISA Coating Buffer, Assay Diluent: 5X Concentrated, Sample Diluent B: 12 mL of a 1X solution per plate, Detection Enzyme: Pre-titrated Streptavidin-HRP, Substrate Solution: Tetramethylbenzidine (TMB) Solution, Certificate of Analysis: Lot-specific instructions for dilution of antibodies and standards, 96 Well Plate: High affinity 96-well plate (included with product Cat. Nos. ending in suffixes -22 and -86)
• Label or Dye: Biotin
• Quantity: 2 x 96 Tests
• Regulatory Status: RUO
• Sample Volume: Serum 50 μL, Supernatant 100 μL
• Storage Requirements: 2-8°C
• Target: IL-25
• Test Time: 1 hr 45 min
• Total Assay Time: 24 hr 30 min

Frequently Asked Questions (FAQs)

What are the critical parameters for successful ELISPOT assay?

A: It is critically important to use a high-affinity binding PVDF membrane plate, rather than regular ELISA plate or nitrocellulose membrane plate. Additionally, highest affinity antibodies for capture work best in this assay. Empirical optimization of conditions such as cell density, stimulus/mitogen, and kinetics is necessary.

I recently bought one of your Ready-Set-Go! ELISA Sets, can I use the antibody pairs to run an ELISPOT?

We do not recommend the use of our ELISA kits for the ELISPOT application. We cannot guarantee the successful use of our ELISA kits in an ELISPOT application.

Do your Ebioscience antibody pairs work in ELISPOT as well?

Most, but not all, of our ELISA antibody pairs work for ELISPOT. We continue to evaluate and optimize our anti-human and anti-mouse cytokine antibody pairs for ELISPOT applications. Please see our protocols page (www.thermofisher.com/us/en/home/references/protocols/cell-and-tissue-analysis/protocols) for a detailed protocol and a reference table with the list of antibody pairs that can be used in ELISPOT.

What are the components of the Ready-Set-Go! ELISPOT Sets?

Capture Antibody: Pre-titrated, Functional Grade (low endotoxin) purified antibody
Detection Antibody: Pre-titrated, biotin-conjugated antibody
10X Sterile Azide-Free Coating Buffer
5X ELISA/ELISPOT Diluent
Detection enzyme: pre-titrated Avidin HRP
Certificate of Analysis: Lot-specific instructions for dilution of antibodies and enzyme

Safety and Handling

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