Invitrogen™ Mouse IL-13 Uncoated ELISA Kit with Plates

Catalog No.	Quantity
50-172-64	10 x 96 Tests
50-172-61	2 x 96 Tests

Catalog No. 50-172-64 Supplier Invitrogen™ Supplier No. 88713786

Description

ELISA

The Mouse Interleukin 13 (IL-13) 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-13. 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.

IL-13 is an immunoregulatory cytokine produced primarily by activated Th2 cells, and is involved in regulating inflammatory and immune responses. IL-13 down-regulates macrophage activity, thereby inhibiting the production of pro-inflammatory cytokines and chemokines, and is involved in allergen-induced asthma through mechanisms independent of IgE and eosinophils. IL-13 also regulates several stages of B cell maturation and differentiation, up-regulates CD23 and MHC class II expression, and promotes IgE isotype switching of B cells. Structurally, IL-13 closely resembles IL-3, IL-4, IL-5 and GM-CSF. The gene encoding IL-13, IL3, IL5, IL4, and CSF2 form a cytokine gene cluster on chromosome 5q, with the IL-13 gene particularly close to IL4.

Specifications

• Accession Number: P20109
• Assay Range: 8-500 pg/mL
• Assay Sensitivity: 8 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
  eBioscience ELISA/ ELISPOT Coating Buffer Powder: This uncoated ELISA kit with plates may contain ELISA/ELISPOT Coating Buffer Powder (Reconstitute to 1L with dH20 and filter (0.22 μM)) or 10X PBS ELISA Coating Buffer (Dilute 1 part 10X Buffer into 9 parts dH20).
  Assay Diluent: 5X concentrated
  Detection enzyme: Pre-titrated Avidin-HRP
  Substrate Solution: Tetramethylbenzidine (TMB) Substrate Solution
  Certificate of Analysis: Lot-specific instructions for dilution of antibodies and standards
  96 Well Plate: Corning Costar 9018 (included with product Cat. Nos. ending in suffixes -22, -44, -76, -86)
• 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: ALRH; ALRH antibody; BHR1; il 13; Il13; Il-13; IL-13; cytokine; ILN; Interleukin; interleukin 13; Interleukin13; interleukin-13; interleukin-13 precursor; L13; MGC116786; MGC116788; MGC116789; NC30; NC30 antibody; P600; P600 antibody; T-cell activation protein P600ALRH; ALRH antibody; BHR1; il 13; Il13; Il-13; IL-13; cytokine; ILN; Interleukin; interleukin 13; Interleukin13; interleukin-13; interleukin-13 precursor; L13; MGC116786; MGC116788; MGC116789; NC30; NC30 antibody; P600; P600 antibody; T-cell activation protein P600; IFG; IFI
• Gene ID (Entrez): 16163
• Gene Symbol: Il13
• Kit Contents: Capture Antibody: Pre-titrated, purified antibody, Detection Antibody: Pre-titrated, HRP-conjugated antibody, Standard: Recombinant protein for generating standard curve and calibrating samples, 10X Coating Buffer: Buffer for plating the Capture Antibody, 5X ELISA/ELISPOT Diluent: Buffer for blocking and diluting the Detection Antibody and Enzyme, Substrate: 1X 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, -76, -86), Purified antibody, Detection Antibody: Pre-titrated, HRP-conjugated antibody, Standard: Recombinant protein for generating standard curve and calibrating samples, 10X Coating Buffer: Buffer for plating the Capture Antibody, 5X ELISA/ELISPOT Diluent: Buffer for blocking and diluting the Detection Antibody and Enzyme, Substrate: 1X 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, -76, -86)
• Label or Dye: Biotin
• Protein Aliases: IL-13, T-cell activation protein P600
• Quantity: 10 x 96 Tests
• Regulatory Status: RUO
• Sample Volume: Serum 100 μL, Supernatant 100 μL
• Storage Requirements: 2-8°C
• Synonym: Interleukin-13, IL13
• Target: IL-13
• 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