Antigen InformationAccession Number:O00585Gene ID:6366Gene Symbols:CCL21SCYA21UNQ784/PRO1600Protein Name & Synonyms:C-C motif chemokine 21 (6Ckine) (Beta-chemokine exodus-2) (Secondary lymphoid-tissue chemokine) (SLC) (Small-inducible cytokine A21)Target Species:HumanAssay FormatSensitivity:3 pg/ml (Anticipated minimum sensitivity. Assay may detect lower levels of antigen.)Number of Targets Detected:1Species Detected:HumanCompatible Sample Types:Cell Culture SupernatantsPlasmaSerumDesign Principle:Sandwich-basedMethod of Detection:qPCRQuantitative/Semi-Quantitative:QuantitativeSolid Support:96-well MicroplateProduct SpecificationsSize:1, 2, or 5 x 96-Well Strip Microplate Kit NEED HELP RUNNING THIS ASSAY? Full testing service for only $5 More per plate REQUEST QUOTE | MORE INFO IntroductionThe RayBio® Immuno Quantitative Enzyme Linked ImumunoSorbent Assay (IQELISA™) is an innovative new assay that combines the specificity and ease of use of an ELISA with the sensitivity of real-time PCR. This results in an assay that is simultaneously familiar and cutting edge and enables the use of lower sample volumes while also providing more sensitivity. The RayBio® Human 6Ckine IQELISA™ Kit is a modified ELISA assay with high sensitivity qPCR readout for the quantitative measurement of Human 6Ckine in serum, plasma, and cell culture supernatants. This assay employs an antibody specific for Human 6Ckine coated on a 96-well PCR plate. Standards and samples are pipetted into the wells and 6Ckine present in a sample is bound to the wells by the immobilized antibody. The wells are washed and a detection affinity molecule is added to the plates. After washing away unbound detection affinity molecule, primers and a PCR master mix are added to the wells and data is collected using qPCR. Ct values obtained from the qPCR are then used to calculate the amount of antigen contained in each sample, where lower Ct values indicate a higher concentration of antigen.Typical DataApplication NotesKit Components6Ckine Microplate (Item A): 96 well PCR plate coated with anti-Human 6CkineWash Buffer I Concentrate (20x) (Item B): 25 ml of 20x concentrated solutionStandards (Item C): 2 vials of recombinant Human 6CkineAssay Diluent A (Item D): 30 ml diluent buffer, 0.09% sodium azide as preservative. For Standard/Sample (serum/plasma) diluentAssay Diluent B (Item E): 15 ml of 5x concentrated buffer. For Standard/Sample (cell culture medium/urine) diluentDetection Affinity Reagent for 6Ckine (Item F): 2 vials of a 4x concentrated solution of anti-Human 6Ckine affinity reagentIQELISA™ Detection Reagent (Item G): 1.4ml of a 10x concentrated stockPrimer Solution (Item I): 1.7 ml vialPCR Master Mix (Item J): 1.2 ml vialPCR Preparation buffer (Item K): 1ml vial of 10x concentrated bufferFinal Wash Buffer (Item L): 10ml vial of 10x concentrated bufferOther Materials RequiredReal-time PCR instrument, Bio-Rad recommendedPrecision pipettes to deliver 2 µl to 1 ml volumesAdjustable 1-25 ml pipettes for reagent preparation100 ml and 1 liter graduated cylindersAbsorbent paperDistilled or deionized waterLog-log graph paper or computer and software for data analysisTubes to prepare standard or sample dilutionsHeating block or water bath capable of 80°CProtocol OutlinePrepare all reagents, samples and standards as instructedAdd 25µl standard or sample to each well. Incubate for 2.5 hours at room temperature or overnight at 4°CAdd 25 µl detection affinity reagent to each well. Incubate 1 hour at room temperatureAdd 25µL of IQELISA™ Detection Reagent to each well. Incubate 1 hourAdd 15µL Primer solution 10µL of PCR master mix to each wellRun real-time PCRStorage/StabilityMay be stored for up to 6 months at 2° to 8°C from the date of shipment. Standard (recombinant protein) should be stored at -20°C or -80°C (recommended at -80°C) after reconstitution. Opened PCR plate or reagents may be stored for up to 1 month at 2° to 8°C. Note: the kit can be used within one year if the whole kit is stored at -20°C. Avoid repeated freeze-thaw cycles.No posts foundWrite A Review