Mouse/Rat RGM-C/Hemojuvelin Quantikine ELISA Kit

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MRGMC0
Mouse/Rat RGM-C/Hemojuvelin ELISA Standard Curve
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Mouse/Rat RGM-C/Hemojuvelin Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (13 uL), EDTA Plasma (13 uL), Heparin Plasma (13 uL)
Sensitivity
4.46 pg/mL
Assay Range
31.3 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Recombinant and natural mouse and rat RGM-C/Hemojuvelin
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
Interference observed with 1 or more available related molecules.

Product Summary

The Quantikine Mouse/Rat RGM-C/Hemojuvelin Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse and rat RGM-C in cell culture supernates, tissue homogenates, serum, and plasma. It contains NS0-expressed recombinant mouse RGM-C and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural RGM-C showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural RGM-C.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of a known concentration were tested in separate assays to determine inter-assay precision. Assays were performed by at least three technicians using two lots of kit components

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 75 244 751 72.8 228 726
Standard Deviation 3.6 6.1 23 5.35 16.6 47.8
CV% 4.8 2.5 3.1 7.3 7.3 6.6

Recovery

The recovery of mouse/rat RGM-C spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 98 85-110
EDTA Plasma (n=4) 100 80-120
Heparin Plasma (n=4) 100 83-116
Serum (n=4) 105 88-120

Linearity

To assess the linearity of the assay, samples containing high concentrations of RGM-C were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Mouse/Rat RGM-C/Hemojuvelin ELISA Linearity

Scientific Data

Mouse/Rat RGM-C/Hemojuvelin ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: RGM-C/Hemojuvelin

The repulsive guidance molecule (RGM) family of molecules contains three uniquely structured GPI-linked glycoproteins that participate in a variety of activities. Termed RGM-A, -B and -C, they are 40-45 kDa in size, show generally nonoverlapping fetal expression patterns, and serve as BMP-4 receptors. RGM-A and -C are plasma membrane bound, while RGM-B is generally microsomal in location. RGM-A and -B are associated with neurons, while RGM-C regulates skeletal muscle and hepatocyte function.

Long Name:
Repulsive Guidance Molecule C
Entrez Gene IDs:
148738 (Human); 69585 (Mouse); 310681 (Rat)
Alternate Names:
DL-M; haemojuvelin; hemochromatosis type 2 (juvenile); Hemojuvelin; HFE2; HFE2AMGC23953; HJV; HJVHemochromatosis type 2 protein; JH; repulsive guidance molecule c; RGMC; RGM-C; RGMCRGM domain family member C
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

FAQs

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