Human soluble cluster of differentiation 14,sCD14 ELISA Kit

Instructions
Code CSB-E13199h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

This ELISA kit is used for quantitative detection of a human soluble cluster of differentiation 14 (sCD14), establishing normal detection range and validation of antibody array results. CD14 is a pattern-recognition receptor and exists in two different forms: GPI-anchored membrane CD14 (mCD14) and liver-derived serum soluble CD14. sCD14 is an acute-phase protein upregulated significantly during sepsis but is also found in normal serum at microgram concentrations. CD14 plays an important role in the phagocytic clearance of apoptotic cells and the reactivation and immune recognition of microbial cell wall components from Gran-positive and Gram-negative bacteria. sCD14 can transfer the LPS to the TLR4/MD-2 complex, eliciting a cascade of inflammatory signaling and TLR4 endocytosis.

Based on the Sandwich-ELISA principle and enzyme-substrate chromogenic reaction property, the users can measure the content of human CD14 in serum, plasma, cell culture supernates, tissue homogenates, or urine. This human CD14 ELISA kit is quality tested with high specificity, excellent sensitivity, precision less than 10%, good linearity, and high recovery. Click the product instructions for more validation information.

Target Name CD14 molecule
Alternative Names CD 14 ELISA Kit; CD_antigen=CD14 ELISA Kit; CD14 ELISA Kit; CD14 antigen ELISA Kit; CD14 molecule ELISA Kit; CD14_HUMAN ELISA Kit; LPS-R ELISA Kit; Mo2 ELISA Kit; Monocyte differentiation antigen CD14 ELISA Kit; Monocyte differentiation antigen CD14 urinary form ELISA Kit; Monocyte differentiation antigen CD14; membrane-bound form ELISA Kit; Myeloid cell specific leucine rich glycoprotein ELISA Kit; Myeloid cell-specific leucine-rich glycoprotein ELISA Kit
Abbreviation CD14
Uniprot No. P08571
Species Homo sapiens (Human)
Sample Types serum, plasma, cell culture supernates, tissue homogenates, urine
Detection Range 0.234 ng/mL-15 ng/mL
Sensitivity 0.229 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Immunology
Assay Principle quantitative
Measurement Sandwich
Precision

 

Intra-assay Precision (Precision within an assay): CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess.

Inter-assay Precision (Precision between assays):CV%<10%

Three samples of known concentration were tested in twenty assays to assess.

 

Linearity

 

To assess the linearity of the assay, samples were spiked with high concentrations of human sCD14 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.

 

Typical Data

 

These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.

 

Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-human CD14 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled CD14 antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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 Q&A
Q:

For item CSB-E13199h,if the kit detects both soluble and membrane bound CD14. Would you have any testing data that addresses this question that you could share?

A:
Thanks for your inquiry.
CD14 exists in two forms, one anchored to the membrane by a glycosylphosphatidylinositol tail (mCD14), the other a soluble form (sCD14).
Soluble CD14 either appears after shedding of mCD14 or is directly secreted from intracellular vesicles.
CSB-E13199h This kit is designed according to: https://www.uniprot.org/uniprot/P08571. It detects soluble form.
For serum and plasma samples, CD14 usually exists in soluble form and the kit detects soluble CD14.
For cell samples, after cellwall breaking, the hydrophobic region of the membrane protein will be closed, and the soluble component can be detected at this time.
We tested serum and plasma samples before and the test value is 3.75--7.5ng/ml under 1:200 dilution.
The final concentration is 750--1500ng/ml.
Pls let me know if you have any further questions. Thank you.

Target Data

Function Coreceptor for bacterial lipopolysaccharide
Gene References into Functions
  1. high plasma levels of sCD14 is associated with measles virus infection. PMID: 29795672
  2. a considerable connection of DEFB1 and TCF7L2 gene polymorphisms with nephrolithiasis PMID: 29959006
  3. CD14 C-(260)T polymorphism is not associated with incidence of acute myocardial infarction in Egyptians who showed elevated serum CD14 levels in PMID: 29874995
  4. This study provides better understanding of the mechanisms and disease susceptibility for MIF and CD14 genetic variants and inflammatory miRNAs networks involved in ankylosing spondylitis and polyarthralgia. PMID: 29863307
  5. Childhood-onset and adult-onset of asthma showed significant difference in allergen sensitivity as well as genetic background with respect to CD14 polymorphism. PMID: 29937881
  6. Renal transplant recipients carrying the CD14-159 TT genotype have significantly higher risk of acute rejection and reduced transplant survival rate than patients with heterozygous or wild-type genotypes. PMID: 28411360
  7. The accumulating evidence does not support an association of CD14 SNP rs2569190 with AR risk. [meta-analysis] PMID: 29687183
  8. It seems that CD14 gene polymorphism might be associated with the risk of CAD, whereas COL4A1 gene polymorphism was not found to confer any risk of CAD PMID: 29299748
  9. higher sCD14 levels in HIV-positive women were associated with a more compromised maternal immunological status and to a lower neonatal birthweight, but not to poorer clinical outcomes in the HIV-exposed children PMID: 29323435
  10. The -221G>C polymorphism of MBL2, the -159C>T polymorphism of CD14 and the TNF-857 polymorphism of TNF-a are risk factors for spinal spinal tuberculosis (TB) and may be involved in the development of spinal TB in the Chinese population. These factors are indicators of susceptibility to spinal TB and require clinical attention. PMID: 29298876
  11. genetic variation of CD14, rs5744455, is related to the susceptibility to laryngeal cancer, providing a theoretical basis for the study of the pathogenesis of laryngeal PMID: 29077168
  12. Calcitriol regulates immune genes CD14 and CD180 to modulate LPS responses in human trophoblasts. PMID: 29089453
  13. The findings suggest that smoking and the presence of TNFalpha-308 GA/AA genotypes may increase the risk for peri-implantitis, while CD14-159 polymorphic CT/TT genotypes decrease the risk. PMID: 28906511
  14. Our results demonstrate that the expression of CD163 and CD206 in monocytes is modulated by LPS in vitro; LPS induces CD163 expression and downregulates the spontaneously increased expression of CD206 PMID: 25914252
  15. may be a simple and easy approach to assess by RT-FCM the reaction between NO and superoxide ion in whole blood monocytes..The no-wash, no-lyse staining protocol with CD45-KO and CD14-PB allows to differentiate clearly and to gate in the monocyte population in near-physiological conditions PMID: 25758468
  16. Association of polymorphic markers of chemokine genes, their receptors, and CD14 gene with coronary atherosclerosis PMID: 29369549
  17. data confirm that engineered human cells expressing TLR4, MD2 and CD14 can respond to CMP with NF-kappaB activation and the response can be influenced by variations in CMP-mannosylation PMID: 29281684
  18. hypothesize that CD14(-159C/T) polymorphic variants might be one of genetic components in the response to attenuated M. bovis BCG bacilli PMID: 29281719
  19. Higher sCD14 levels were found in twins with schizophrenia or bipolar disorder compared to their not affected co-twins, and higher sCD14-levels were associated with psychotic symptoms. PMID: 28039552
  20. novel observation that sCD14 compared with lipopolysaccharide binding protein, offers a preferred target to ameliorate TLR especially TLR4-induced inflammation and insulin resistance in human obesity and metabolic syndrome PMID: 26880233
  21. For asthma, there was suggestive evidence of a gene-by-environment interaction for the CD14 variant rs2569190 (Interaction P = 0.16) but not for the TLR4 variants rs4986790 and rs4986791. PMID: 27977294
  22. the simultaneous detection of both plasma sCD14 and IL-6 is a promising diagnostic approach to identify active pulmonary tuberculosis, and further, measurement of TNF-alpha and sCD163 can identify the most severe cases of tuberculosis. PMID: 27591510
  23. the methylation levels in the promoter region of CD14 gene were higher in the Finnish compared to Russian Karelian children PMID: 27434019
  24. Anti-apoA-1 IgG are independent predictors of nonfatal incident coronary artery disease in the general population. The strength of this association is dependent on a functional polymorphism of the CD14 receptor gene, a finding suggesting a gene-autoantibody interaction for the development of CAD. PMID: 29074586
  25. CD14 haplotypes and sCD14 are important mediators of lung function among those with COPD in this occupationally-exposed population. PMID: 28302109
  26. IL-1beta-primed dendritic cells express high levels of CD14 that mediate IL-17 production through direct interaction with T cells. PMID: 27550748
  27. The distribution of the CD14 rs2569190 genotypes and allele frequencies showed significant differences between the test and control groups. Prevalence of the polymorphic TT genotype and T allele of the CD14 rs2569190 were significantly increased in GCP patients when compared with healthy controls PMID: 28829191
  28. Presepsin is significantly higher in preterm infants with early-onset sepsis compared with uninfected infants. PMID: 27925621
  29. Neovascularized atherosclerotic lesions selectively associate with lower blood levels of CD14+ and CD14(high)CD16- monocytes independently of systemic inflammatory activity. PMID: 27751505
  30. CXCR7 mediates CD14(+)CD16(+) monocyte transmigration across the blood brain barrier, and is a potential therapeutic target for neuro AIDS. PMID: 28754798
  31. META-ANALYSIS: association between the -159C/T polymorphism in the promoter region of the CD14 gene and sepsis PMID: 28122493
  32. In LPS-stimulated HEK293 cells with low CD14 and high TLR4, no accumulation of PI(4,5)P2 occurred. With an increasing amount of CD14 and decrease of TLR4, 2 peaks of PI(4,5)P2 appeared, approaching those found in LPS-stimulated cells expressing CD14 alone. This suggests that LPS-induced accumulation of PI(4,5)P2 that maximizes TLR4 signaling is controlled by CD14, whereas TLR4 can fine tune this via PI(4,5)P2 turnover. PMID: 27418354
  33. Intermediate CD14++CD16+monocytes might be closely related to the pathogenesis of atrial fibrillation and reflect functional remodelling of the left atrium. PMID: 26826137
  34. In 121 primary sclerosing cholangitis patients, the CD14 -260C>T genotype was associated with development of dominant bile duct strictures (P = 0.02). In 365 liver transplantation patients, TT carriers (4.1%) were protected against the formation of nonanastomotic biliary strictures versus CC/CT patients (12.6%; P = 0.01). PMID: 26970220
  35. response of hPdLSCs to bacterial LPS is strongly augmented by sCD14 PMID: 27504628
  36. analysis of plasma CXCL10, sCD163 and sCD14 in virological suppression and risk of cardiovascular disease PMID: 27355513
  37. TGM2 has a role in macrophage differentiation via mechanisms involving CD14 and SR-AI receptors. PMID: 27378395
  38. we found that higher percentages of circulating CD14+CD204+, CD14+CD163+CD204+ M2-like monocytes were significantly associated with TNM stage, lymph node metastasis, and histological differentiation. PMID: 28639912
  39. these results indicate that CD14 is a co-receptor of TLR4 in the S100A9-induced cytokine response. PMID: 27228163
  40. our data showed the contribution on the TLR4+896A/G and CD14-159C/T polymorphism-related immune dysfunction including increased non-classical (inflammatory) monocyte proportion-related LPS hyper-inflammatory response and decreased classical (phagocytic) monocyte proportion-related impaired phagocytosis in febrile acute de-compensated cirrhotic patients complicated with severe sepsis. PMID: 27861595
  41. Plasma presepsin concentrations are associated with the severity of sepsis and its outcome. PMID: 28099244
  42. Presepsin is a valuable new biomarker for defining severe infections in cirrhosis, proving same efficacy as procalcitonin. However, it is not a useful marker of short-term mortality. PMID: 27895404
  43. Patients with CD14 C (-159) T gene polymorphism, a co-receptor of TLR4, have an increased risk of NAFLD development. PMID: 27895422
  44. Single nucleotide polymorphism CD14-159 C/T was linked to greater Atopic dermatitis risk at 2 to 3 years of age. PMID: 27274543
  45. While there was no association with any respiratory phenotype (as determined by symptoms), the CD14 CT/TT genotype appeared to be protective to increased exposure to NO2 and NO PMID: 27142357
  46. CD14 expression is significantly upregulated in human masticatory mucosa during wound healing PMID: 28005267
  47. we report that exogenous CnB is taken up by cells in a time- and concentration-dependent manner via clathrin-dependent receptor-mediated internalization. Our findings further confirm that uptake is mediated by the TLR4/MD2 complex together with the co-receptor CD14 PMID: 27090571
  48. Cord blood sCD14 may be a useful biomarker for predicting infant wheeze and prolonged cough by 1 year of age. PMID: 27144407
  49. Presepsin is a valuable biomarker to detect patients with acute pyelonephritis PMID: 27889429
  50. diagnostic accuracy of presepsin (sCD14-ST) as a biomarker of infection and sepsis PMID: 27823951

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Subcellular Location Cell membrane, Lipid-anchor, GPI-anchor, Secreted, Membrane raft, Golgi apparatus
Tissue Specificity Detected on macrophages (at protein level) (PubMed:1698311). Expressed strongly on the surface of monocytes and weakly on the surface of granulocytes; also expressed by most tissue macrophages.
Database Links

HGNC: 1628

OMIM: 158120

KEGG: hsa:929

STRING: 9606.ENSP00000304236

UniGene: Hs.163867

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