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Guinea Pig TGF-β1(Transforming Growth Factor Beta 1) ELISA Kit

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SKU:
GPFI00051

Description

Product name

Guinea Pig TGF-β1(Transforming Growth Factor Beta 1) ELISA Kit

Catalogue No.

GPFI00051

Detection method

Sandwich ELISA, Double Antibody

Size

96T

Range

31.25-2000pg/ml

Sensitivity

< 18.75pg/ml

Storage

4'C for 6 months

Recovery

Matrices listed below were spiked with certain level of and the recovery rates were calculated by comparing the measured value to the expected amount of in samples.

Matrix Recovery range(%) Average(%)
serum(n=5) 85-103 92
EDTA plasma(n=5) 85-101 96
heparin plasma(n=5) 90-105 96
Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 88-92% 95-102% 86-102% 87-104%
EDTA plasma(n=5) 85-93% 85-97% 83-100% 93-100%
heparin plasma(n=5) 87-99% 87-99% 80-100% 81-97%
CV(%)

Intra-Assay: CV<8%
Inter-Assay: CV<10%

Note

For Research Use Only

Datasheets

ELISA Kit Technical ManualELISA Kit Technical ManualMSDS

Protocol

Sandwich ELISA Protocol

The below protocol is a sample protocol for a sandwich ELISA using a biotinylated detection antibody and streptavidin-HRP. Sandwich ELISAs allow for the detection and quantification of an analyte in a sample by using known analyte concentrations as standards and plotting absorbance of known concentrations vs known standard concentrations. This allows the researcher to calculate the amount of analyte present in their sample. This can be very useful when looking for increases in protein concentration, phosphorylation of proteins or decreases on protein concentrations.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at room temperature (37 °C). When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

Sandwich Protocol

ELISA Kit Technical Manual

Procedure:

1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 15-30 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.
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Additional Information

Reactivity:
Guinea pig
ELISA Type:
Sandwich
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