Human Attractin Quantikine ELISA Kit

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DATRN0
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Human Attractin Standard Curve
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Human Attractin 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 (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Saliva (10 uL), Urine (10 uL)
Sensitivity
0.217 ng/mL
Assay Range
0.8 - 50 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine)
Specificity
Natural and recombinant human Attractin.
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.

Product Summary

The Quantikine Human Attractin Immunoassay is a 4.5 hour solid phase ELISA designed to measure Attractin levels in cell culture supernates, serum, plasma, saliva, and urine. It contains CHO cell-expressed recombinant human Attractin and antibodies raised against the recombinant protein. Results obtained for naturally occurring human Attractin showed linear curves that were parallel to the standard curves obtained using the Quantikine Human Attractin Immunoassay standards. These results indicate that this kit can be used to determine relative mass values for natural human Attractin.

Precision

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

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 4.16 12.7 26.2 4.61 13.7 27.5
Standard Deviation 0.132 0.324 0.893 0.498 1.1 1.54
CV% 3.2 2.6 3.4 10.8 8 5.6

Recovery

The recovery of human Attractin spiked to levels throughout the range of the assay was evaluated

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 91 87-97

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Attractin were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Attractin ELISA Linearity

Product Datasheets

Preparation and Storage

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

Background: Attractin

Attractin/ATRN, also known as DPPT-L, is an approximately 200 kDa transmembrane glycoprotein that shows dipeptidyl peptidase activity similar to DPPIV/CD26. Attractin is involved in a variety of processes including monocyte-T cell adhesion, axon myelination, melanocyte pigment synthesis, and energy homeostasis. Attractin is transiently upregulated during T cell activation before expression switches to the 175 kDa secreted isoform which is released into the circulation. Soluble Attractin is preferentially expressed by leukocytes and differentiating neurons. It blocks neurite formation and is elevated in the CSF of astrocytoma patients.

Entrez Gene IDs:
8455 (Human); 11990 (Mouse); 83526 (Rat)
Alternate Names:
ATRN; Attractin; DPPT-L; KIAA0548; Mahogany Protein; MGC126754; MGCA
⚠ 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. 100 µL Assay Diluent
  4.   Add 100 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 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 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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