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Human MBL Quantikine ELISA Kit

R&D Systems | Catalog # DMBL00

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL)

Sensitivity

0.029 ng/mL

Assay Range

0.156-10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Human MBL Quantikine ELISA Kit

The Quantikine Human MBL immunoassay is a 4.5 hour solid phase ELISA designed to measure human MBL in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human MBL and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human MBL showed dose-response curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values of naturally occurring human MBL.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Oligomeric forms of natural and recombinant human MBL

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

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 0.91 2.05 4.06 1.02 2.06 4.14
Standard Deviation 0.03 0.07 0.22 0.08 0.14 0.29
CV% 3.3 3.4 5.4 7.8 6.8 7.0

Recovery for Human MBL Quantikine ELISA Kit

The recovery of MBL spiked to various levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=6) 113 106-117

Linearity

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

Human MBL ELISA Linearity

Scientific Data Images for Human MBL Quantikine ELISA Kit

Human MBL ELISA Standard Curve

Human MBL ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped with polar packs. 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: MBL

Mannose/mannan-binding lectin (MBL) is a secreted glycoprotein in the collectin family of pattern-recognition molecules. It contains a cysteine-rich region, a collagen-like segment, and a C-type lectin domain that binds to neutral bacterial carbohydrates. The collagen-like region mediates MBL association into disulfide-stabilized trimers which further associate into complexes containing three or four copies of the basic trimer. MBL is secreted by hepatocytes and opsonizes bacteria through interactions with microbial carbohydrates. MBL multimers can associate with the serine proteases MASP-1, -2, and -3 and promote their cleavage of Complement Component C3. Proteolytic cleavage of C3 triggers activation of the complement system and formation of the membrane attack complex, leading to destruction of opsonized bacteria. Opsonized apoptotic material is cleared by the scavenger receptor CD91.

Long Name

Mannose Binding Lectin

Alternate Names

COLEC1, Collectin-1, MBL2, MBL2D, MBP1

Entrez Gene IDs

4153 (Human)

Gene Symbol

MBL2

Additional MBL Products

Product Documents for Human MBL 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 Human MBL 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.

Citations for Human MBL Quantikine ELISA Kit

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Protocols

View specific protocols for Human MBL Quantikine ELISA Kit (DMBL00):

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
  3.   Add 50 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 room temperature for 2 hours on a horizontal orbital microplate shaker.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 100 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 100 µL Substrate Solution
  8.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 100 µL Stop Solution
  9.   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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FAQs

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