This kit is based on Double antibody-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with anti REG1alpha antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with REG1alpha bound to precoated antibody. Wash unbound components and add HRP-Streptavidin Conjugate (SABC). Wash unbound components again and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of REG1alpha in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Alternative Names
Lithostathine-1-alpha ELISA Kit, Islet cells regeneration factor ELISA Kit, ICRF ELISA Kit, Islet of Langerhans regenerating protein ELISA Kit, REG ELISA Kit, Pancreatic stone protein ELISA Kit, PSP ELISA Kit, Pancreatic thread protein ELISA Kit, PTP ELISA Kit, Regenerating islet-derived protein 1-alpha ELISA Kit, REG-1-alpha ELISA Kit, Regenerating protein I alpha ELISA Kit, REG1A ELISA Kit, PSPS ELISA Kit, PSPS1 ELISA Kit, REG ELISA Kit
Detection Method
Double Antibody
Kit Components
Kit Components
Item
Size(48T)
Size(96T)
Storage Condition for Opened Kit
E001
ELISA Microplate(Dismountable)
8x6
8x12
Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E002
Lyophilized Standard
1vial
2vial
Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E003
Biotin-labeled Antibody(Concentrated, 100X)
60ul
120ul
2-8°C (Avoid Direct Light)
E034
HRP-Streptavidin Conjugate(SABC, 100X)
60ul
120ul
E024
TMB Substrate
5ml
10ml
E039
Sample Dilution Buffer
10ml
20ml
2-8°C
E040
Antibody Dilution Buffer
5ml
10ml
E049
SABC Dilution Buffer
5ml
10ml
E026
Stop Solution
5ml
10ml
E038
Wash Buffer(Concentrated, 25X)
15ml
30ml
E006
Plate Sealer
3 pieces
5 pieces
E007
Product Description
1 copy
1 copy
Recovery
Add a certain amount of REG1alpha into the sample. Calculate the recovery by comparing the measured value with the expected amount of REG1alpha in the sample.
Sample Type
Recovery Range(%)
Average(%)
serum(n=10)
88-104
95
EDTA plasma(n=10)
90-94
92
Heparin plasma(n=10)
86-105
95
Linearity
Dilute the sample with a certain amount of REG1alpha at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type
1:2
1:4
1:8
serum(n=10)
92-97%
90-102%
88-105%
EDTA plasma(n=10)
84-100%
90-97%
85-91%
Heparin plasma(n=10)
82-97%
80-94%
83-100%
Specificity
Specifically binds with REG1alpha , no obvious cross reaction with other analogues.
Standard Curve
This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.) Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.
Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate. Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.
Item
Intra-assay Precision
Inter-assay Precision
Sample
1
2
3
1
2
3
n
20
20
20
20
20
20
Mean (pg/ml)
126.9
492.3
2018
125.7
507.7
2030
Standard deviation
5.8
25.7
106.35
6.01
25.94
97.64
CV(%)
4.57
5.22
5.27
4.78
5.11
4.81
Background
REG1α
Stability
Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.
ELISA kit(n=5)
37°C for 1 month
2-8°C for 6 months
Average(%)
80
95-100
Notes
For Research Use Only
Usage Notes
Step 1: Take out the required plate wells, add 50ul Cap/Det Ab into each well, then add 50ul Standard or Sample into individual well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure thorough mixing then static incubate for 60 minutes at 37°C.
Washing: Wash the plate twice without immersion.
Step 2: Add 100ul HRP-Streptavidin (orange) into each well, seal the plate and static incubate for 30 minutes at 37°C.
Washing: Wash the plate five times without immersion.
Step 3: Add 90ul TMB substrate solution, seal the plate and static incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
Step 4: Add 50ul stop solution. Read at 450nm immediately and calculate.
Antigen
REG1alpha
UniProt ID
P05451
Applications
ELISA
Applications Notes
REG1alpha ELISA Kit allows for the in vitro quantitative determination of REG1alpha concentrations in serum, plasma, tissue homogenates and other biological fluids.
Storage
2-8°C(Sealed), Don't cryopreserve.
Reviews of Human REG1alpha (RegeneRating Islet Derived Protein 1 Alpha) ELISA Kit