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Human Properdin / Complement Factor P ELISA Kit

SKU:
HUFI00956
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P27918
Sensitivity:
0.938ng/ml
Range:
1.563-100ng/ml
ELISA Type:
Sandwich
Synonyms:
CFP, Complement Factor P, Properdin, PFC, BFD, complement factor properdin, PFCcomplement
Reactivity:
Human
Research Area:
Immunology
€499
Frequently bought together:

Description

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Human Properdin/Complement Factor P ELISA Kit

The Human Properdin Complement Factor P ELISA Kit is a powerful tool for the precise measurement of properdin levels in human samples such as serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit delivers dependable and consistent results, making it perfect for various research purposes.Properdin is a key component of the complement system, playing a crucial role in enhancing immune responses and inflammation.

Dysregulation of properdin levels has been implicated in various diseases, including autoimmune disorders and infections, making it a valuable biomarker for studying these conditions and exploring potential treatments.Get accurate and reliable data on properdin levels with the Human Properdin Complement Factor P ELISA Kit, and unlock new insights into the role of properdin in health and disease.

Product Name:Human Properdin / Complement Factor P ELISA Kit
Product Code:HUFI00956
Size:96 Assays
Alias:CFP, Complement Factor P, Properdin, PFC, BFD, complement factor properdin, PFCcomplement
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human CFP concentrations in serum plasma and other biological fluids.
Sensitivity:0.938ng/ml
Range:1.563-100ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human CFP and the recovery rates were calculated by comparing the measured value to the expected amount of Human CFP in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)89-10397
EDTA plasma(n=5)85-10596
UFH plasma(n=5)87-10597
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human CFP and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)87-102%85-103%87-99%
EDTA plasma(n=5)84-95%83-101%85-97%
UFH plasma(n=5)84-100%82-98%80-97%
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotP27918
UniProt Protein Function:CFP: A positive regulator of the alternate pathway of complement. It binds to and stabilizes the C3- and C5-convertase enzyme complexes. Defects in CFP are the cause of properdin deficiency (PFD). PFD results in higher susceptibility to bacterial infections; especially to meningococcal infections. Three phenotypes have been reported: complete deficiency (type I), incomplete deficiency (type II), and dysfunction of properdin (type III).
UniProt Protein Details:Protein type:Secreted, signal peptide; Secreted

Chromosomal Location of Human Ortholog: Xp11.4

Cellular Component: extracellular matrix; extracellular space; endoplasmic reticulum lumen; extracellular region

Biological Process: protein amino acid O-linked glycosylation; cellular protein metabolic process; defense response to bacterium; complement activation, alternative pathway; regulation of complement activation; innate immune response; immune response; post-translational protein modification; complement activation

Disease: Properdin Deficiency, X-linked

NCBI Summary:This gene encodes a plasma glycoprotein that positively regulates the alternative complement pathway of the innate immune system. This protein binds to many microbial surfaces and apoptotic cells and stabilizes the C3- and C5-convertase enzyme complexes in a feedback loop that ultimately leads to formation of the membrane attack complex and lysis of the target cell. Mutations in this gene result in two forms of properdin deficiency, which results in high susceptibility to meningococcal infections. Multiple alternatively spliced variants, encoding the same protein, have been identified.[provided by RefSeq, Feb 2009]
UniProt Code:P27918
NCBI GenInfo Identifier:464473
NCBI Gene ID:5199
NCBI Accession:P27918.2
UniProt Secondary Accession:P27918,O15134, O15135, O15136, O75826,
UniProt Related Accession:P27918
Molecular Weight:469
NCBI Full Name:Properdin
NCBI Synonym Full Names:complement factor properdin
NCBI Official Symbol:CFP
NCBI Official Synonym Symbols:BFD; PFC; PFD; PROPERDIN
NCBI Protein Information:properdin; complement factor P; properdin P factor, complement
UniProt Protein Name:Properdin
UniProt Synonym Protein Names:Complement factor P
Protein Family:Properdin
UniProt Gene Name:CFP
UniProt Entry Name:PROP_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample TypeProtocol
SerumIf using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

PlasmaCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal FluidCollect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatantCollect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysatesSolubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenatesThe preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysatesRinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast MilkCollect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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