Human Calbindin D Quantikine ELISA Kit

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DCALD0
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Human Calbindin D Standard Curve
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Human Calbindin D 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
Tissue Lysates (10 uL), Urine (25 uL)
Sensitivity
0.015 ng/mL
Assay Range
0.1 - 5 ng/mL
Specificity
Natural and recombinant human Calbindin D.
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 Calbindin D Immunoassay is a 4.5 hour solid phase ELISA designed to measure human Calbindin D in tissue lysates and urine. It contains E. coli-expressed recombinant human Calbindin D and antibodies raised against the recombinant factor. Natural human Calbindin D showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative levels of natural human Calbindin D.

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

Tissue Lysates, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 0.511 1.65 2.84 0.548 1.57 2.96
Standard Deviation 0.026 0.04 0.072 0.038 0.075 0.15
CV% 5.1 2.4 2.5 6.9 4.8 5.1

Recovery

The recovery of human Calbindin D spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Urine (n=4) 93 89-98

Linearity

To assess the linearity of the assay, samples containing high concentrations of human Calbindin D were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Human Calbindin D ELISA Linearity

Product Datasheets

Preparation and Storage

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

Background: Calbindin D

Calbindin D is a highly conserved vitamin D-dependent cytoplasmic protein in the EF-hand calcium binding protein family that includes calmodulin, parvalbumin, troponin C, and S100. Calbindin-D is widely expressed and regulates calcium homeostasis. Calbindin-D protects neurons from excitotoxic and apoptotic cell death by buffering excess calcium.

Long Name:
Vitamin D-dependent Calcium-Binding Protein
Entrez Gene IDs:
795 (Human); 12309 (Mouse); 24249 (Rat)
Alternate Names:
CABP; CABP1; CABP28K; CABP9K; Calbindin 3; Calbindin D; Calbindin-D; Calbindin-D9K; S100G
⚠ 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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ELISA Controls

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