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Intrinsic Mouse Erythroferrone™ ELISA Kit

$788.00

Species: Mouse Analyte: Erythroferrone
Range: 0.06 – 4 ng/ml Sensitivity: 0.014 ng/ml
SKU: ERF-200 Category: Tags: , ,
Catalog NumberERF-200
Quantity96 Wells
ApplicationELISA, EIA, RUO
MethodSandwich
Antibody TypeMonoclonal
Target SpeciesMouse
AnalyteErythroferrone
Assay time2h 45m
Detection methodColorimetric
Sample typeSerum
Sample volume required per well10ul
Detection range0.0625-4ng/ml
Sensitivity0.014ng/ml
Regulatory statusResearch Use Only ELISA
Storage requirements4C

Typical Data

Hepcidin StandardsConcentration (ng/mL)Absorbance (450nm)
42.196
21.321
10.727
0.50.41
0.250.243
0.1250.146
0.0630.10
0.0010.056

Precision

Intra assay PrecisionInter assay Precision
Sample123123
n161616161616
Mean (ng/ml)0.56.215.30.56.014.5
Standard Deviation0.030.260.560.030.350.63
CV(%)5.64.23.77.25.74.3

Linearity

Serum
1:2Range (%)90-111
1:4Range (%)97-123
1:8Range (%)96-117
1:16

Recovery

Sample TypeRange(%)Average Recovery(%)
Serum89-109101

Description

The Intrinsic Mouse ERFETM ELISA is a test kit designed for quantification of mouse Erythroferrone in serum. This kit does not contain mercaptoethanol, azide- or mercury-based preservatives.

Detection of mouse erythroferrone is based on the double antibody sandwich ELISA method. The 96-well microplate assay plate is pre-coated with anti-mouse erythroferrone polyclonal antibody. Mouse serum samples, standards, or controls are mixed with buffer and incubated in the assay plate. Mouse erythroferrone present in the sample binds to the immobilized antibody on the assay plate. The plate is then washed to remove unbound mouse erythroferrone. A 2nd anti-mouse erythroferrone antibody conjugated with biotin is added and followed with HRP-SA conjugate which binds to biotinylated anti-mouse erythroferrone. Optical density (OD) is measured by the addition of TMB, the reaction produces a blue color and development is halted with the addition of the stop solution. The OD is read at 450nm on a microplate reader and a standard curve is prepared by plotting the log concentration of the standard curve versus its corresponding OD. The intensity of the color is proportional to the concentration of ERFE in the sample. Total assay run time is less than 3 hours.

Note:
Based on our studies, serum ERFE concentrations in healthy mice (C57BL/6) were essentially undetectable (below the limit of detection). In a mouse model of ineffective erythropoiesis (beta-thalassemia), serum ERFE concentrations ranged from 1-2 ng/ml.

ERF-200 Product Insert

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General Information

Catalog number ERF-200
Quantity 96 Wells
Application ELISA, EIA, RUO
Method Sandwich
Antibody type Polyclonal
Target species Mouse
Analyte Erythroferrone
Assay time 2h 45m
Detection method Colormetric
Sample type Serum
Sample volume required per well 10 ul
Detection range 0.0625 – 4.0 ng/ML
Sensitivity 0.014 ng/mL
Regulatory Status Research use only ELISA
Storage requirements 2-8 c
Shelf Life 3 Months from Kit Assembly

Typical data

Erythroferrone Standard Concentration (ng/ML) Absorbance (450nm)
1 4 2.196
2 2 1.321
3 1 0.727
4 0.5 0.410
5 0.25 0.243
6 0.125 0.146
7 0.063 0.100
8 0.001 0.056

Precision

Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 16 16 16 16 16 16
Mean(ng/ml) 0.5 6.2 15.3 0.5 6.0 14.5
Standard deviation 0.03 0.26 0.56 0.03 0.35 0.63
CV (%) 5.6 4.2 3.7 7.2 5.7 4.3

Linearity

Serum
1:2 Range (%) 90-111
Average (%) 99
1:4 Range (%) 97-123
Average (%) 106
1:8 Range (%) 96-117
Average (%) 109

Recovery

Sample Type Range (%) Average Recovery (%)
Serum 89-109 101

Additional information

Weight 16 oz
Dimensions 7.0 × 7 × 4.0 in