Mouse/Rat RGM-A Quantikine ELISA Kit

R&D Systems | Catalog # MRGMA0

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Tissue Homogenates (50 µL), Serum (13 µL), EDTA Plasma (13 µL), Heparin Plasma (13 µL)

Sensitivity

0.019 ng/mL

Assay Range

0.078-5 ng/mL (Cell Culture Supernates, Tissue Homogenates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Mouse/Rat RGM-A Quantikine ELISA Kit

The Quantikine Mouse/Rat RGM-A Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse and rat RGM-A in cell culture supernates, tissue homogenates, serum, and plasma. It contains NS0-expressed recombinant mouse RGM-A and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural RGM-A 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-A.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Mouse, Rat

Specificity

Natural and recombinant mouse and rat RGM-A

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

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 known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum, Tissue Homogenates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/mL) 0.190 0.742 2.12 0.195 0.538 2.07
Standard Deviation 0.010 0.018 0.087 0.015 0.029 0.147
CV% 5.3 2.4 4.1 7.7 5.4 7.1

Recovery for Mouse/Rat RGM-A Quantikine ELISA Kit

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

Sample Type Average % Recovery Range %
Mouse Cell Culture Supernates (n=4) 101 91-112
Mouse EDTA Plasma (n=4) 106 91-119
Mouse Heparin Plasma (n=4) 102 95-117
Mouse Serum (n=4) 113 103-120
Mouse Tissue Homogenates (n=4) 100 93-106
Rat Cell Culture Supernates (n=4) 100 90-113
Rat EDTA Plasma (n=4) 107 103-115
Rat Heparin Plasma (n=4) 99 91-106
Rat Serum (n=4) 107 94-119

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of RGM-A were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Mouse/Rat RGM-A ELISA Linearity
Mouse/Rat RGM-A ELISA Linearity

Scientific Data Images for Mouse/Rat RGM-A Quantikine ELISA Kit

Mouse/Rat RGM-A ELISA Standard Curve

Mouse/Rat RGM-A ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: RGM-A

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 A

Alternate Names

RGMA

Entrez Gene IDs

56963 (Human); 244058 (Mouse)

Gene Symbol

RGMA

Additional RGM-A Products

Product Documents for Mouse/Rat RGM-A Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse/Rat RGM-A Quantikine ELISA Kit

For research use only

⚠ 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.

Related Research Areas

Citations for Mouse/Rat RGM-A Quantikine ELISA Kit

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Protocols

View specific protocols for Mouse/Rat RGM-A Quantikine ELISA Kit (MRGMA0):

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.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 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.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. 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.

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