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Human Pro-Cathepsin B ELISA Kit - Quantikine

Catalog #: DCATB0
9 Citations 1 Review Datasheet / COA / SDS
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DCATB0
Control Products Available
QC58: Quantikine Immunoassay Control Set 426 Human Pro-Cathepsin B
Human Pro-Cathepsin B ELISA Serum/Plasma Standard Curve
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Citations (9)
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Reviews (1)
Summary Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

Human Pro-Cathepsin B 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), Cell Lysates (25 uL), Serum (10 uL), Heparin Plasma (10 uL), Saliva (10 uL), Urine (25 uL)
Sensitivity
0.079 ng/mL
Assay Range
0.2 - 10 ng/mL (Cell Culture Supernates, Cell Lysates, Serum, Heparin Plasma, Saliva, Urine)
Specificity
Natural and recombinant human Pro-Cathepsin B
Cross-reactivity
Cross-reactivity observed with 1 or more 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 Pro-Cathepsin B Immunoassay is a 4.5 hour solid phase ELISA designed to measure the pro-form of cathepsin B in cell culture supernates, cell lysates, serum, plasma, saliva, and urine. It contains NS0-expressed recombinant human Pro-Cathepsin B and antibodies raised against the recombinant factor. Natural human Pro-Cathepsin B showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards, indicating that this kit can be used to determine relative levels of natural human Pro-Cathepsin B.

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, Cell Lysates, Saliva, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 1.08 2.39 4.5 1.16 2.58 4.91
Standard Deviation 0.05 0.11 0.23 0.1 0.16 0.3
CV% 4.6 4.6 5.1 8.6 6.2 6.1

Serum, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 1.32 2.93 5.78 1.48 3.18 5.95
Standard Deviation 0.08 0.15 0.29 0.13 0.24 0.42
CV% 6.1 5.1 5 8.8 7.5 7.1

Recovery

The recovery of Pro-Cathepsin B spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 106 98-115

Linearity

To assess the linearity of the assay, samples containing high concentrations of Pro-Cathepsin B were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Pro-Cathepsin B ELISA Linearity

Scientific Data

N/A Human Pro-Cathepsin B ELISA Serum/Plasma Standard Curve View Larger

Human Pro-Cathepsin B ELISA Serum/Plasma Standard Curve

N/A Human Pro-Cathepsin B ELISA Cell Culture Supernate/Cell Lysate/Saliva/Urine Standard Curve View Larger

Human Pro-Cathepsin B ELISA Cell Culture Supernate/Cell Lysate/Saliva/Urine 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: Cathepsin B

Cathepsin B is the first described member of the family of lysosomal cysteine proteases. Cathepsin B possesses both endopeptidase and exopeptidase activities, in the latter case acting as a peptidyl-dipeptidase. It is known to process a number of proteins, including pro and active caspases, prorenin and SLPI. Cathepsin B is synthesized as a preproenzyme. Following removal of the signal peptide, the inactive proenzyme undergoes further modifications including removal of the pro region to result in the active enzyme.

Entrez Gene IDs:
1508 (Human); 13030 (Mouse)
Alternate Names:
APP secretase; APPS; Cathepsin B; Cathepsin B1; CPSBamyloid precursor protein secretase; CTSB; cysteine protease; EC 3.4.22; EC 3.4.22.1
⚠ 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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at 2-8 °C for 2 hours.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at 2-8 °C for 2 hours.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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.

Citations for Human Pro-Cathepsin B Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

9 Citations: Showing 1 - 9
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  1. Dysregulation of macrophage-secreted cathepsin B contributes to HIV-1-linked neuronal apoptosis.
    Authors: Rodriguez-Franco EJ, Cantres-Rosario YM, Plaud-Valentin M, Romeu R, Rodriguez Y, Skolasky R, Melendez V, Cadilla CL, Melendez LM
    PLoS ONE, 2012-05-31;7(5):e36571.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. A possible contribution of altered cathepsin B expression to the development of skin sclerosis and vasculopathy in systemic sclerosis.
    Authors: Noda S, Asano Y, Akamata K, Aozasa N, Taniguchi T, Takahashi T, Ichimura Y, Toyama T, Sumida H, Yanaba K, Tada Y, Sugaya M, Kadono T, Sato S
    PLoS ONE, 2012-02-23;7(2):e32272.
    Species: Human
    Sample Types: Serum
  3. Identification of candidate nasopharyngeal carcinoma serum biomarkers by cancer cell secretome and tissue transcriptome analysis: potential usage of cystatin A for predicting nodal stage and poor prognosis.
    Authors: Chang KP, Wu CC, Chen HC
    Proteomics, 2010-07-01;10(14):2644-60.
    Species: Human
    Sample Types: Serum
  4. Decreased lubricin concentrations and markers of joint inflammation in the synovial fluid of patients with anterior cruciate ligament injury.
    Authors: Elsaid KA, Fleming BC, Oksendahl HL, Machan JT, Fadale PD, Hulstyn MJ, Shalvoy R, Jay GD
    Arthritis Rheum., 2008-06-01;58(6):1707-15.
    Species: Human
    Sample Types: Synovial Fluid
  5. Imbalanced serum concentration between cathepsin B and cystatin C in patients with pelvic inflammatory disease.
    Authors: Tsai HT, Wang PH, Tee YT, Lin LY, Hsieh YS, Yang SF
    Fertil. Steril., 2008-03-07;91(2):549-55.
    Species: Human
    Sample Types: Serum
  6. Mesenchymal stem cells support migration, extracellular matrix invasion, proliferation, and survival of endothelial cells in vitro.
    Authors: Potapova IA, Gaudette GR, Brink PR, Robinson RB, Rosen MR, Cohen IS, Doronin SV
    Stem Cells, 2007-03-29;25(7):1761-8.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Generation of endostatin by matrix metalloproteinase and cathepsin from human limbocorneal epithelial cells cultivated on amniotic membrane.
    Authors: Ma DH, Yao JY, Kuo MT, See LC, Lin KY, Chen SC, Chen JK, Chao AS, Wang SF, Lin KK
    Invest. Ophthalmol. Vis. Sci., 2007-02-01;48(2):644-51.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Acidic extracellular pH promotes experimental metastasis of human melanoma cells in athymic nude mice.
    Authors: Rofstad EK, Mathiesen B, Kindem K, Galappathi K
    Cancer Res., 2006-07-01;66(13):6699-707.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. Serum level of cathepsin B and its density in men with prostate cancer as novel markers of disease progression.
    Authors: Miyake H, Hara I, Eto H
    Anticancer Res., 2004-07-01;24(4):2573-7.
    Species: Human
    Sample Types: Serum

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Human Pro-Cathepsin B Quantikine ELISA Kit
By Anonymous on 08/29/2018
Sample Tested: EDTA Plasma,Serum