This assay employs the quantitative sandwich enzyme immunoassay technique (See Fig. 1). A monoclonal antibody specific for human TGF-β1 was pre-coated onto a microplate. Standards and samples are pipetted into the wells, and incubated with biotin-conjugated detection antibody (Detection A) specific for human TGF-β1. After incubation, the plate wells are aspirated without washing. Then, incubate with Streptavidin- HRP (Detection B). Following a wash to remove any unbound antibody and samples, an ultra-sensitive TMB substrate solution is added to the wells for color development. The color intensity is in proportion to the amount of TGF-β1 bound in the initial step. The intensity of the color is measured by plate read at 450 nm.
Typical Data
This standard curve(R2=0.9995) is provided for demonstration only. A standard curve should be generated for each set of samples assayed. Fig. 3 is an example of typical Data.
Sensitivity
The minimum detectable dose (MOD) of human TGF-β1 is typically 10 pg/ml.
The Intra-assay CV is 3.79% the Inter-assay CV is ˂10%.
Specificity
This assay recognizes natural and recombinant human TGF-β1.
Simple Procedures
Simple Standard Preparation
Accurate Standard Curve
Kit Content and Storage Conditions
PART
PART#
DESCRIPTION
STORAGE OF OPENED/ RECONSTITUTED
Human TGF-β1 Microplate
TBS3085A
96 well polystyrene microplate (12 strips of 8 wells) coated with a polyclonal antibody specific for human TGF-β1.
Return unused wells to the foil pouch. Reseal along the entire edge of the zip-seal. May be stored for up to 1 month at 2-8 °C.
Human TGF-β1 Standard
TBS3085B
50 µl of Recombinant human TGF-β1 protein (100ng/mL).
Aliquot and store at -20 ℃ for up to 1 month in a manual defrost the freezer. Avoid repeated freeze-thaw cycles.
Detection A
TBS3085C
2.1 ml of Biotin-Human TGF-β1 antibody.
May be stored for up to
3 months at 2-8 °C.*
Detection B
TBS3085D
300 µl of Streptavidin-HRP.
Assay Diluent
TBS3085E
25 ml of a buffered protein base with preservatives.
Wash Buffer
TBS3000W
12 ml of concentrated solution (10x).
TMB Substrate
TBS3000T
12 ml of ultra-sensitive TMB substrate.
Stop Solution
TBS3000S
6 ml of 2 N sulfuric acid.
Store the unopened kit at 2-8 ℃. Do not use past kit expiration dates.
The kit contains sufficient materials to run an ELISA on one 96 well plate.
