Invitrogen™ Human/Mouse TGF beta-1 Uncoated ELISA Kit

Catalog No.	Quantity
50-174-93	10 x 96 Tests
50-174-91	20 x 96 Tests

Catalog No. 50-174-93 Supplier Invitrogen™ Supplier No. 88835088

Description

ELISA

The Human/Mouse Transforming Growth Factor beta 1 (TGFβ1) Uncoated ELISA Kit contains pre-matched antibody pairs, and reagents for performing quantitative enzyme linked immunosorbent assays (ELISA) to detect and quantify protein levels of human/mouse TGFβ1. 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.

TGF beta-1 (TGFB1, Transforming Growth Factor Beta 1) is a polypeptide member of the transforming growth factor beta superfamily of cytokines, found almost ubiquitously in tissues. Transforming growth factor (TGF)-b is stored in the extracellular matrix as a latent complex with its pro-domain. Activation of TGF beta-1 requires the binding of aV integrin to an RGD sequence in the prodomain and exertion of force on this domain, which is held in the extracellular matrix by latent TGF-b binding proteins. Latent forms are complexes of TGF beta-1, an amino-terminal portion of the TGF-beta precursor, designated TGF-LAP (TGF-latency associated peptide), and a specific binding protein, known as LTBP. TGF beta-1 helps regulates proliferation, differentiation, adhesion, migration in many cell types. Many cells have TGF beta receptors, and the protein positively and negatively regulates many other growth factors. TGF beta-1 is cleaved into a latency-associated peptide and a mature TGF beta-1 peptide, and is found in either a latent form composed of a TGFB1 homodimer, a LAP homodimer, and a latent TGFB1-binding protein, or in an active form composed of a TGF beta-1 homodimer. The mature peptide may also form heterodimers with other TGF beta family members. The gene for TGF beta-1 is frequently upregulated in tumor cells, and mutations in this gene result in Camurati-Engelmann disease and cystic fibrosis.

Specifications

• Accession Number: P01137, P04202
• Assay Range: 15.6-1000 pg/mL
• Assay Sensitivity: 15.6 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
  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, -76, and -86)
• Conjugate: HRP
• Immunoassay Kit Format: Uncoated ELISA Kit
• Product Type: ELISA
• Sample Type: Plasma, Serum, Supernatant
• Target Species: Human, Mouse
• For Use With (Application): ELISA
• For Use With (Equipment): Absorbance Microplate Reader
• Gene Alias: CED; cytokine; DPD1; LAP; Latency-associated peptide; prepro-transforming growth factor beta-1; regulatory protein; TGF b; TGF b 1; TGF beta; tgf beta 1; tgf beta1; TGF beta-1; TGF beta-1 protein; TGF β; TGF β 1; TGF β1; Tgfb; TGFB1; TGF-B1; Tgfb-1; TGFbeta; TGF-beta 1; TGF-beta 1 protein; TGFbeta1; TGF-beta1; TGF-beta-1; TGFβ1; Transforming growth factor; transforming growth factor b1; transforming growth factor beta; transforming growth factor beta 1; transforming growth factor beta-1; Transforming growth factor beta-1 proprotein; transforming growth factor beta-1 proprotein; transforming growth factor beta-1; transforming growth factor β1; transforming growth factor, beta 1; transforming growth factor, beta 1 (Camurati-Engelmann disease); transforming growth factor, beta-1; transforming growth factor-beta 1; transforming growth factor-beta I; transforming growth factor-beta-1; transforming growth factor-beta-1 precursor; unnamed protein productCED; cytokine; DPD1; LAP; Latency-associated peptide; prepro-transforming growth factor beta-1; regulatory protein; TGF b; TGF b 1; TGF beta; tgf beta 1; tgf beta1; TGF beta-1; TGF beta-1 protein; TGF β; TGF β 1; TGF β1; Tgfb; TGFB1; TGF-B1; Tgfb-1; TGFbeta; TGF-beta 1; TGF-beta 1 protein; TGFbeta1; TGF-beta1; TGF-beta-1; TGFβ1; Transforming growth factor; transforming growth factor b1; transforming growth factor beta; transforming growth factor beta 1; transforming growth factor beta-1; Transforming growth factor beta-1 proprotein; transforming growth factor beta-1 proprotein; transforming growth factor beta-1; transforming growth factor β1; transforming growth factor, beta 1; transforming growth factor, beta 1 (Camurati-Engelmann disease); transforming growth factor, beta-1; transforming growth factor-beta 1; transforming growth factor-beta I; transforming growth factor-beta-1; transforming growth factor-beta-1 precursor; unnamed protein product; DIF-alpha; TNFA; TNFSF2; TNLG1F; TNF
• Gene ID (Entrez): 21803, 7040
• Gene Symbol: Tgfb1
• 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, 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 Plate: High affinity 96-well plate (included with product Cat. Nos. ending in suffixes -22, -76, -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, 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 Plate: High affinity 96-well plate (included with product Cat. Nos. ending in suffixes -22, -76, -86)
• Label or Dye: Biotin
• Protein Aliases: TGF-beta-1, LAP
• Quantity: 10 x 96 Tests
• Regulatory Status: RUO
• Sample Volume: Plasma 20 μL, Serum 20 μL, Supernatant 100 μL
• Storage Requirements: 2-8°C
• Synonym: Transforming Growth Factor beta 1, TGF-b1
• Target: TGF beta-1
• 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