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Regenerating Islet-Derived Family, Member 4 (REG4) ELISA Kit
|2 references available|
|Price||862.63 $ Plus shipping costs $45.00|
|Availability||Will be delivered in 9 to 11 Business Days|
|Method type||Sandwich ELISA|
|Alternative name||regenerating gene IV (REG-4)|
|Cross-Reactivity (Details)||No significant cross-reactivity or interference between Human REG4 and analogues was observed.|
|Sample Type||Serum, Plasma|
|Sample Volume||50 - 100 µL|
|Assay Time||1 - 4.5 h|
|Plate||Pre-coated 8 x 12 strip plate|
1. Assay plate (12 x 8 coated Microwells). Quantity: 1(96 wells)
2. Standard (Freeze dried). Quantity: 2
3. Biotin-antibody (100 x concentrate) Quantity: 1 x 120 µl
3.HRP-avidin (100 x concentrate). Quantity: 1 x 120 µl
4. Biotin-antibody Diluent. Quantity: 1 x 10 ml
5. HRP-avidin Diluent. Quantity: 1 x 10 ml
6. Sample Diluent. Quantity: 1 x 20 ml
7. Wash Buffer (25 x concentrate). Quantity: 1 x 20 ml
8. TMB Substrate. Quantity: 1 x 10 ml
9. Stop Solution. Quantity: 1 x 10 ml
10. Adhesive Strip (For 96 wells). Quantity: 4
11. Instruction manual
|Description||Synonyms: GISP, REG-IV, RELP, OTTHUMP00000014031|gastrointestinal secretory protein|regenerating gene type IV|
|Characteristics||This immunoassay kit allows for the in vitro quantitative determination of human REG-4 concentrations in cell culture supernates, serum, plasma and other biological fluids.|
|Specificity||This assay has high sensitivity and excellent specificity for detection of Human REG4.|
|Sensitivity||The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined the mean O.D value of 20 replicates of the zero standard added by their three standard deviations.|
|Minimum Detection Limit||0.078 ng/mL|
|Detection Range||0.312 ng/mL - 20 ng/mL|
|Principle||This assay employs the quantitative sandwich enzyme immunoassay technique.|
|Protocol||Antibody specific for REG4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any REG4 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for REG4 is added to the wells. After washing, avidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of REG4 bound in the initial step. The color development is stopped and the intensity of the color is measured.|
Bring all reagents and samples to room temperature before use. Centrifuge the sample again after thawing before the assay. It is recommended that all samples and standards be assayed in duplicate.
|Calculation of Results||Using the professional soft 'Curve Expert 1.3' to make a standard curve is recommended, which can be downloaded from our web. Average the duplicate readings for each standard and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the REG4 concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.|
|Application Notes||Detection wavelength: 450 nm|
|Material not included||
1. Microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
2. An incubator which can provide stable incubation conditions up to 37° C +/- 0.5° C.
3. Squirt bottle, manifold dispenser, or automated microplate washer.
4. Absorbent paper for blotting the microtiter plate.
5. 100 mL and 500 mL graduated cylinders.
6. Deionized or distilled water.
7. Pipettes and pipette tips.
8. Test tubes for dilution.
Unopened: Store at 2 - 8° C. Do not use past kit expiration date.
Opened: May be stored for up to 1 month at 2 - 8° C. Try to keep coated assay plate in a sealed aluminium foil bag, and avoid the damp.
|Restrictions||For Research Use only|
Zheng, Xu, Yu et al.: "The role of Reg IV gene and its encoding product in gastric carcinogenesis." in: Human pathology, Vol. 41, Issue 1, pp. 59-69, 2009 (PubMed).
Tao, He, Ma et al.: "Evaluation of REG4 for early diagnosis and prognosis of gastric cancer." in: Human pathology, 2011 (PubMed). Method employed by authors: ELISA (Sample species: Human).