Rat Hepcidin (Hepc) (Sandwich ELISA) ELISA Kit

Mfr.No.

HEB979Ra

Sensitivity

87.3pg/mL

Detection Range

246.9-20000pg/mL

Reactivity

Rat

Assay Duration

Assay Type

Sandwich

Description

This assay has high sensitivity and excellent specificity for detection of High Sensitive Hepcidin (Hepc).
No significant cross-reactivity or interference between High Sensitive Hepcidin (Hepc) and analogues was observed.

Alternative Names

HAMP; HFE2B; PLTR; LEAP1; Hepcidin Antimicrobial Peptide; Liver-expressed antimicrobial peptide 1; Putative liver tumor regressor

Detection Method

Colormetric

Kit Components

Reagents	Quantity	Reagents	Quantity
Pre-coated, ready to use 96-well strip plate	1	Plate sealer for 96 wells	4
Standard	2	Standard Diluent	1×20mL
Detection Reagent A	1×120µL	Assay Diluent A	1×12mL
Detection Reagent B	1×120µL	Assay Diluent B	1×12mL
TMB Substrate	1×9mL	Stop Solution	1×6mL
Wash Buffer (30 × concentrate)	1×20mL	Instruction manual	1

Recovery

Matrices listed below were spiked with certain level of recombinant High Sensitive Hepcidin (Hepc) and the recovery rates were calculated by comparing the measured value to the expected amount of High Sensitive Hepcidin (Hepc) in samples.

Matrix	Recovery range (%)	Average(%)
serum(n=5)	81-101	88
EDTA plasma(n=5)	93-104	101
heparin plasma(n=5)	79-89	83

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of High Sensitive Hepcidin (Hepc) 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)	83-97%	78-93%	93-101%	80-101%
EDTA plasma(n=5)	86-101%	85-101%	79-105%	89-97%
heparin plasma(n=5)	85-95%	96-104%	78-97%	82-103%

Features

Detects protein (low-target protein, high-sensitivity)

Species

Rat

Precautions

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Hepcidin (Hepc) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Hepcidin (Hepc) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: C<10%
Inter-Assay: C<12%

Notes

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to High Sensitive Hepcidin (Hepc) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled High Sensitive Hepcidin (Hepc) and unlabeled High Sensitive Hepcidin (Hepc) (Standards or samples) with the pre-coated antibody specific to High Sensitive Hepcidin (Hepc). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of High Sensitive Hepcidin (Hepc) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of High Sensitive Hepcidin (Hepc) in the sample.

Usage Notes

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
And then add 50µL prepared Detection Reagent A immediately.
Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

Gene ID	84604
UniProt ID	Q99MH3

Applications	ELISA
Research Area	Metabolic pathway

Storage	4°C for 1 month/ -20°C for long-term(within shelf life)
Shelf Life	one year

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SKU	Supplier No.	Size	Your price
BHE10104810	HEB979Ra	96T*1	$768	Add to Cart
BHE10104810	HEB979Ra	96T*5	$3456	Add to Cart
BHE10104810	HEB979Ra	96T*10	$6528	Add to Cart

High Sensitive ELISA Kit for Hepcidin (Hepc) (Rat)

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