Human proteoglycan 4(PRG4) ELISA Kit

Code CSB-E14124h
Size 96T,5×96T,10×96T
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Product Details

Alternative Names
PRG4 ELISA Kit; MSF ELISA Kit; SZP ELISA Kit; Proteoglycan 4 ELISA Kit; Lubricin ELISA Kit; Megakaryocyte-stimulating factor ELISA Kit; Superficial zone proteoglycan) [Cleaved into: Proteoglycan 4 C-terminal part] ELISA Kit
Abbreviation
PRG4
Uniprot No.
Species
Homo sapiens (Human)
Sample Types
serum, plasma and tissue homogenates.
Detection Range
0.78 ng/ml-50 ng/ml.
Sensitivity
0.195 ng/ml
Assay Time
1-5h
Sample Volume
50-100ul
Detection Wavelength
450 nm
Research Area
Others
Assay Principle
quantitative
Measurement
Sandwich
Materials provided
    • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human PRG4 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 PRG4 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-antibodyDiluent (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
Storage
Store at 2-8°C. Please refer to protocol.
Lead Time
3-5 working days after you place the order, and it takes another 3-5 days for delivery via DHL or FedEx
Description

The Human proteoglycan 4 (PRG4) ELISA kit is suitable for quantitative measurement of PRG4 levels in the serum, plasma, or tissue homogenates. It is not intended for diagnostic use and only used for scientific research in humans. The kit has undergone rigorous quality control in multiple parameters, including sensitivity, specificity, precision, linearity, recovery, and inter-batch difference. Refer to the product instructions for more details.

This assay employs the quantitative sandwich enzyme immunoassay technique, in which PRG4 in the samples or standards are sandwiched between pre-coated PRG4 antibody and Biotin-conjugated PRG4 antibody. HRP-avidin is then added to the wells. Following a wash to remove any unbound reagent, the TMB substrate solution is added to the wells and color develops in proportion to the amount of PRG4 bound in the initial step. The color development is stopped upon adding the stop solution, and the intensity of the color is measured at 450 nm via a microplate reader. The levels of PRG4 in the samples can be determined by referring to the O.D. (optical density) of the samples to the standard curve.

PRG4, also called lubricin, is a mucinous glycoprotein secreted by synovial fibroblasts and superficial zone chondrocytes. It is a major component of synovial fluid (SF) and therefore plays an important role in joint lubrication and synovial homeostasis. PRG4, which is naturally present at the human cornea-eyelid interface, has been proved to be involved in the reduction of the friction between the cornea and the conjunctiva. Marwa Qadri etc. revealed that PRG4 is an endogenous antifibrotic modulator in the joint and its effect on myofibroblast formation is partially mediated by CD44, but CD44 is not required to demonstrate an antifibrotic effect in vivo. Loss of PRG4 function has been linked to camptodactyly-arthropathy-coxa vara-pericarditis syndrome (CACP), an arthritis-like autosomal recessive disorder.

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Target Background

Function
(From Uniprot)
Plays a role in boundary lubrication within articulating joints. Prevents protein deposition onto cartilage from synovial fluid by controlling adhesion-dependent synovial growth and inhibiting the adhesion of synovial cells to the cartilage surface.; Isoform F plays a role as a growth factor acting on the primitive cells of both hematopoietic and endothelial cell lineages.
Gene References into Functions
  1. Inflammatory Biomarker Profiling in Total Joint Arthroplasty and Its Relevance to Circulating Levels of Lubricin, a Novel Proteoglycan. PMID: 29683034
  2. Intra-articular injection of human PRG4 in vivo in Prg4-/- mice prevented caspase-3 activation in superficial zone chondrocytes and was associated with a modest decrease in whole joint friction. PMID: 28604608
  3. PRG4 plays an important anti-inflammatory role in regulating osteoarthritis synoviocyte proliferation. PMID: 28482921
  4. IL6 and PRG4 represent potential novel tissue biomarkers of disease severity and prognosis in conjunctival fibrosis after glaucoma surgery. PMID: 28975281
  5. adult talar cartilage increases both PRG4 release and biosynthetic activity as immediate cellular response to injury PMID: 27551813
  6. Double knockdown of PRG4 and IL-24 did not inhibit myxoid liposarcoma (MLS)- cell growth, and single knockdown of PRG4 remarkably increased IL-24 expression. These results suggest that the growth inhibitory effect of PRG4 knockdown is caused by induction of IL-24 expression, and PRG4 may contribute to maintain MLS cell growth through repression of IL-24 expression. PMID: 28192118
  7. lubricin expression may typify adaptive and neoplastic changes along a pathway toward fibroblast-like synoviocytes PMID: 26924731
  8. PRG4 binds to TLR2 and TLR4 and this binding mediates a novel anti-inflammatory role for PRG4. PMID: 26643105
  9. Cartilage derived from MSCs expressed lubricin protein both in vitro and in vivo PMID: 26867127
  10. no synovial fluid level differences detected between healthy knees and injured knees PMID: 26037740
  11. The finding that rhPRG4 can increase the viscosity of low concentration HA solutions suggests that supplementation with rhPRG4 may help mitigate the loss in synovial fluid viscosity experienced with decreased HA concentration in osteoarthritis. PMID: 25818000
  12. PRG4 is a novel putative ligand for CD44 and may control synoviocyte overgrowth in inflammatory arthropathies via a CD44-mediated mechanism. PMID: 25708025
  13. The O-glycomap of lubricin, a novel mucin responsible for joint lubrication, has been identified by site-specific glycopeptide analysis. PMID: 25187573
  14. Lubricin (Prg4) plays a role in preventing damage to the superficial zone and preservation of chondrocytes. [Review] PMID: 25172828
  15. 5 novel PRG4 mutations and the first case of CACP syndrome resulting from uniparental disomy of chromosome 1. PMID: 23756439
  16. We speculate that an important role of lubricin in mediating interactions at the cartilage surface is to attach to the cartilage surface and provide a protective coating that maintains the contacting surfaces in a sterically repulsive state. PMID: 24406099
  17. Lubricin is transcribed, translated, and expressed by ocular surface epithelia. Lubricin presence significantly reduces friction between the cornea and conjunctiva. PMID: 23599181
  18. The objective was to evaluate the presence and distribution of the lubricating and anti-adhesion glycoprotein lubricin and cells containing the contractile isoform smooth muscle alpha-actin (SMA) in pseudomembranes around loose hip prostheses. PMID: 23174700
  19. the identification of a novel null mutation in PRG4 confirming the genetic homogeneity of Camptodactyly-arthropathy-coxa vara-pericarditis syndrome . PMID: 22678705
  20. Production and accumulation of the superficial zone protein (SZP), also known as lubricin, by the surface zone is a characteristic feature of articular cartilage. PMID: 22490392
  21. Lubricin in human breast tissue expander capsules PMID: 22865664
  22. lubricin is able to bind to PMN via an L-selectin-dependent and -independent manner and may play a role in PMN-mediated inflammation. PMID: 22930755
  23. lubricin is expressed in the TMJ disc bilaminar zone; lubricin may have a role in normal disc posterior attachment physiology through the prevention of cellular adhesion as well as providing lubrication during normal bilaminar zone function PMID: 21955422
  24. Lubricin is expressed in chondrocytes derived from osteoarthritic cartilage encapsulated in poly (ethylene glycol) diacrylate scaffold. PMID: 22073377
  25. The surface layer of lubricin coating torn edges of anterior cruciate ligaments and menisci may interfere with the integrative healing process needed for repair. PMID: 21647956
  26. We described a 2-bp novel deletion mutation in PRG4 gene in a Pakistani family with camptodactyly-arthropathy-coxa-vara-pericarditis syndrome PMID: 21565623
  27. O-linked oligosaccharides NeuAc alpha2-3Gal beta1-3GalNAc and NeuAc alpha2-3Gal beta1-3(NeuAc alpha2-6)GalNAc were the dominating structures on lubricin. The latter was more prevalent in rheumatoid arthritis, indicating that sialylation is up-regulated. PMID: 20443780
  28. HAPO enhanced total adherence of HUVEC in a concentration-dependent manner. PMID: 19900364
  29. Mrna present in tendons from tennis elbow. PRG4 may also be expressed as alternatively spliced form lacking exons which encode part of the N-terminal matrix-binding and cell-proliferative domain. PMID: 12475643
  30. Megakaryocyte stimulating factor (msf) is linked to prosthetic loosening. PMID: 12783322
  31. data suggest that HAPO is a novel growth factor acting on the primitive cells of both hematopoietic and endothelial cell lineages [HAPO] PMID: 14976050
  32. hemangiopoietin is encoded by HAPO, also known as CACP, MSF, SZP, and PRG4 [editorial] PMID: 15710563
  33. Results describe the production of antibodies against human lubricin to determine the consequence of disease-causing mutations at the protein level and to study the protein's normal post-translational processing. PMID: 16000300
  34. PRG4 mutations may have a role camptodactyly-arthropathy-coxa vara-pericarditis in Saudi families PMID: 16429407
  35. In the human knee, cartilaginous deposits and osteoarthritic cartilage contained PRG4 in patients with advanced knee osteoarthritis. PMID: 17343281
  36. Lubricin provides synovial fluid with an ability to dissipate strain energy induced by mammalian locomotion, which is a chondroprotective feature that is distinct from boundary lubrication. PMID: 17404241
  37. synovial fluid lubricin concentrations were significantly reduced at an early stage following anterior cruciate ligament injury when compared with those in the contralateral joint. PMID: 18512776
  38. These findings demonstrate that HAPO induces endothelial cell proliferation through the PI-3K/Akt pathway. PMID: 18769058
  39. findings point to two distinct mechanisms by which rh-lubricin lubricates, one mechanism involving lubricin bound to the tissue surface and the other involving lubricin in solution PMID: 19058183

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Involvement in disease
Camptodactyly-arthropathy-coxa vara-pericarditis syndrome (CACP)
Subcellular Location
Secreted.
Tissue Specificity
Highly expressed in synovial tissue, cartilage and liver and weakly in heart and lung. Isoform B is expressed in kidney, lung, liver, heart and brain. Isoform C and isoform D are widely expressed.
Database Links

HGNC: 9364

OMIM: 208250

KEGG: hsa:10216

STRING: 9606.ENSP00000399679

UniGene: Hs.647723

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