Collagen Detection

Collagenisthemostabundantproteinfamilythatformsconnectivetissuesandplayscriticalrolesintheinflammatoryprocess.Inordertostudythephysiologicalandpathologicalturnoverofconnectivetissuesandforvariousresearchpurposes,ChondrexprovidesuniquekitsandreagentsfordeterminingvariousspeciesoftypeIandtypeIIcollagen.EachkitisdesignedtoquantifytheamountofnativecollageninculturedtissuesortissuespecimensbyELISA. Toaidresearchersindecidingwhichkittouse,ChondrexprovidesaCollagenDetectionKitSelectionFlowchartbelow.

TypeICollagenDetectionKit

TypeIICollagenDetectionKit

HydroxyprolineAssayKit

ProteinasesForCollagenSolubilization

Semi-QuantitativeTissueTotalCollagenDetectionKit

SiriusRedTotalCollagenDetectionKit

NOTE:Solubilizationofcollagenfromsamplesisrequiredforassayingcollagen.Additionalsuppliessuchasguanidinehydrochloride,pepsin(catalog#6011),elsatase(catalog#30047)etcarenecessaryforprocessingsamples.

Pleasecontactcustomerservicetoobtain"TipsforCollagenSolubilization".

BriefKitDescriptions

TypeICollagenDetectionKits

TypeIcollagenisafibrilarcollagenconsistingoftwoidenticalalpha1(I)chainsandonealpha2(I)chain.Itisthemostabundantcollagentypeandisfoundinmostconnectivetissuessuchasskin,bone,tendon,ligament,andheart.Manytissueswiththeexceptionofbone,whichconsistsoftypeIcollagen,containheterotypicfibrilswithtwoormoredistinctcollagentypesincoexistence.Ingeneral,typeIcollagenisdominantandtypeIIIcollagenisaminorcomponent.Chondrexprovides5typesoftypeIcollagenassaykitfordeterminingbovine,human,mouse,porcineandrattypeIcollagen.

TypeIICollagenDetectionKit

TypeIIcollagenisuniqueamongthecollagenfamily,anditstissuedistributionislimitedtoavasculartissuessuchascartilageandthevitreousbodyoftheeyes.SincetypeIIcollagencaninducearthritisinexperimentalanimals,autoimmunitytotypeIIcollagenissuspectedinthepathogenesisofcertainautoimmunediseasesinhumanssuchasrheumatoidarthritis,eyediseasesassociatedwithrheumatoidarthritis,andrelapsingpolychondritis,whichaffectsspecifictissuescontainingtypeIIcollagen.TheTypeIICollagenDetectionKit(catalog#6018)isdesignedtoquantifytheamountofnativetypeIIcollagenfromvariousspecies(suchashuman,monkey,porcine,bovine,rat,mouse,rabbit,equine,andchick)inculturedtissuesortissuespecimensbyELISA.

Semi-QuantitativeTotalTissueCollagenDetectionKit(SiriusRed/FastGreenCollagenStainingKit)

Thesemi-quantitativetotaltissuecollagendetectionkit(SiriusRed/FastGreencollagenstainingkit:catalog#9046)providesasimplequantitativemicro-assaytoolfordeterminingthetotalamountofcollagenandnon-collagenousproteinsinculturedcelllayersandtissuesectionsbydifferentialstainingwithtwodyes,siriusredandfastgreen.SiriusRedbindstoalltypesofcollagen,whereasfastgreenstainsnon-collagenousproteins.Thismethodhasbeenappliedtothemeasurementofcollagencontentsinvarioustissues.

Thesedyescanbeeasilyextractedfromstainedtissues,andtheamounts(micrograms)ofcollagenandnon-collagenousproteinsineachsectioncanbecalculatedbasedonOD540(siriusred)andOD605(fastgreen).Theassaysensitivityismorethansufficienttodeterminecollagenandnon-collagenousproteinsin10-20micrometertissuesectionspreparedforgeneralhistologicalstudies.Theassaysensitivityforcollagenismorethan3micrograms/section,andnon-collagenousproteinismorethan50microgrampersection.

SiriusRedTotalCollagenDetectionKit 

Siriusredisauniquedyewhichspecificallybindstothe[Gly-X-Y]nhelicalstructureonfibrillarcollagen(typeI-V).SiriusRedTotalCollagenDetectionKit(Catalog#9062)employssiriusredasadetectionreagentdeterminingcollgenconcentrationinsamples.However,thiskitdoesnotdiscriminatebetweencollagenspeciesandtypes.