Mouse C-Ros Oncogene 1, Receptor Tyrosine Kinase (ROS1) ELISA Kit

two product lines: Traditional ELISA Kit and Ready-to-Use ELISA Kit.

	Traditional ELISA Kit	Ready-to-Use ELISA KIT
Product name:	Mouse C-Ros Oncogene 1, Receptor Tyrosine Kinase (ROS1) ELISA Kit
Method:	Sandwich
Synonyms:	MCF3; ROS; c-Ros-1; v-Ros Avian UR2 Sarcoma Virus Oncogene Homolog 1; Proto-oncogene tyrosine-protein kinase ROS; Receptor tyrosine kinase c-ros oncogene 1
Detection range:	78.1-5000pg/mL
Target Protein:	ROS1
Size:	96T/48T
Quality guarantee period:	for 12 months, 16 months
Catalog number:	DL-ROS1-Mu (traditional)	(ready-to-use)
Assay length	1-4.5Hours	1-3.5Hours
Advantages:	Competitive price. High sensitivity. High stability. 12 months shelf life.	Pre-diluted Detection Reagent A and B Reduction in the number of steps when conducting the test All the reagents can be stored at -20℃ Faster reaction compare to other brands 16 months shelf life
Instruction Manual

Mouse C-Ros Oncogene 1, Receptor Tyrosine Kinase (ROS1) ELISA Kit

1. Overview

Other names：AHNAKRS; AHNAK Nucleoprotein; Neuroblast differentiation-associated protein AHNAK

Function：May be required for neuronal cell differentiation.

Sequence：
MEKEETTREL  LLPNWQGSGS  HGLTIAQRDD  GVFVQEVTQN  SPAARTGVVK
EGDQIVGATI  YFDNLQSGEV  TQLLNTMGHH  TVGLKLHRKG  DRSPEPGQTW
TREVFSSCSS  EVVLSGDDEE  YQRIYTTKIK  PRLKSEDGVE  GDLGETQSRT
ITVTRRVTAY  TVDVTGREGA  KDIDISSPEF  KIKIPRHELT  EISNVDVETQ
SGKTVIRLPS  GSGAASPTGS  AVDIRAGAIS  ASGPELQGAG  HSKLQVTMPG
IKVGGSGVNV  NAKGLDLGGR  GGVQVPAVDI  SSSLGGRAVE  VQGPSLESGD
HGKIKFPTMK  VPKFGVSTGR  EGQTPKAGLR  VSAPEVSVGH  KGGKPGLTIQ
APQLEVSVPS  ANIEGLEGKL  KGPQITGPSL  EGDLGLKGAK  PQGHIGVDAS
APQIGGSITG  PSVEVQAPDI  DVQGPGSKLN  VPKMKVPKFS  VSGAKGEETG
IDVTLPTGEV  TVPGVSGDVS  LPEIATGGLE  GKMKGTKVKT  PEMIIQKPKI
SMQDVDLSLG  SPKLKGDIKV  SAPGVQGDVK  GPQVALKGSR  VDIETPNLEG
TLTGPRLGSP  SGKTGTCRIS  MSEVDLNVAA  PKVKGGVDVT  LPRVEGKVKV
PEVDVRGPKV  DVSAPDVEAH  GPEWNLKMPK  MKMPTFSTPG  AKGEGPDVHM
TLPKGDISIS  GPKVNVEAPD  VNLEGLGGKL  KGPDVKLPDM  SVKTPKISMP
DVDLHVKGTK  VKGEYDVTVP  KLEGELKGPK  VDIDAPDVDV  HGPDWHLKMP
KMKMPKFSVP  GFKAEGPEVD  VNLPKADVDI  SGPKIDVTAP  DVSIEEPEGK
LKGPKFKMPE  MNIKVPKISM  PDVDLHLKGP  NVKGEYDVTM  PKVESEIKVP
DVELKSAKMD  IDVPDVEVQG  PDWHLKMPKM  KMPKFSMPGF  KAEGPEVDVN
LPKADVDISG  PKVGVEVPDV  NIEGPEGKLK  GPKFKMPEMN  IKAPKISMPD
VDLHMKGPKV  KGEYDMTVPK  LEGDLKGPKV  DVSAPDVEMQ  GPDWNLKMPK
IKMPKFSMPS  LKGEGPEFDV  NLSKANVDIS  APKVDTNAPD  LSLEGPEGKL
KGPKFKMPEM  HFRAPKMSLP  DVDLDLKGPK  MKGNVDISAP  KIEGEMQVPD
VDIRGPKVDI  KAPDVEGQGL  DWSLKIPKMK  MPKFSMPSLK  GEGPEVDVNL
PKADVVVSGP  KVDIEAPDVS  LEGPEGKLKG  PKFKMPEMHF  KTPKISMPDV
DLHLKGPKVK  GDVDVSVPKV  EGEMKVPDVE  IKGPKMDIDA  PDVEVQGPDW
HLKMPKMKMP  KFSMPGFKGE  GREVDVNLPK  ADIDVSGPKV  DVEVPDVSLE
GPEGKLKGPK  FKMPEMHFKA  PKISMPDVDL  NLKGPKLKGD  VDVSLPEVEG
EMKVPDVDIK  GPKVDISAPD  VDVHGPDWHL  KMPKVKMPKF  SMPGFKGEGP
EVDVKLPKAD  VDVSGPKMDA  EVPDVNIEGP  DAKLKGPKFK  MPEMSIKPQK
ISIPDVGLHL  KGPKMKGDYD  VTVPKVEGEI  KAPDVDIKGP  KVDINAPDVE
VHGPDWHLKM  PKVKMPKFSM  PGFKGEGPEV  DMNLPKADLG  VSGPKVDIDV
PDVNLEAPEG  KLKGPKFKMP  SMNIQTHKIS  MPDVGLNLKA  PKLKTDVDVS
LPKVEGDLKG  PEIDVKAPKM  DVNVGDIDIE  GPEGKLKGPK  FKMPEMHFKA
PKISMPDVDL  HLKGPKVKGD  MDVSVPKVEG  EMKVPDVDIK  GPKVDIDAPD
VEVHDPDWHL  KMPKMKMPKF  SMPGFKAEGP  EVDVNLPKAD  IDVSGPSVDT
DAPDLDIEGP  EGKLKGSKFK  MPKLNIKAPK  VSMPDVDLNL  KGPKLKGEID
ASVPELEGDL  RGPQVDVKGP  FVEAEVPDVD  LECPDAKLKG  PKFKMPEMHF
KAPKISMPDV  DLHLKGPKVK  GDADVSVPKL  EGDLTGPSVG  VEVPDVELEC
PDAKLKGPKF  KMPDMHFKAP  KISMPDVDLH  LKGPKVKGDV  DVSVPKLEGD
LTGPSVGVEV  PDVELECPDA  KLKGPKFKMP  EMHFKTPKIS  MPDVDLHLKG
PKVKGDMDVS  VPKVEGEMKV  PDVDIKGPKM  DIDAPDVDVH  GPDWHLKMPK
MKMPKFSMPG  FKAEGPEVDV  NLPKADVVVS  GPKVDVEVPD  VSLEGPEGKL
KGPKLKMPEM  HFKAPKISMP  DVDLHLKGPK  VKGDVDVSLP  KLEGDLTGPS
VDVEVPDVEL  ECPDAKLKGP  KFKMPEMHFK  TPKISMPDVN  LNLKGPKVKG
DMDVSVPKVE  GEMKVPDVDI  RGPKVDIDAP  DVDVHGPDWH  LKMPKMKMPK
FSMPGFKGEG  PEVDVNLPKA  DVDVSGPKVD  VEVPDVSLEG  PEGKLKGPKF
KMPEMHFKTP  KISMPDVDFN  LKGPKIKGDV  DVSAPKLEGE  LKGPELDVKG
PKLDADMPEV  AVEGPNGKWK  TPKFKMPDMH  FKAPKISMPD  LDLHLKSPKA
KGEVDVDVPK  LEGDLKGPHV  DVSGPDIDIE  GPEGKLKGPK  FKMPDMHFKA
PNISMPDVDL  NLKGPKIKGD  VDVSVPEVEG  KLEVPDMNIR  GPKVDVNAPD
VQAPDWHLKM  PKMKMPKFSM  PGFKAEGPEV  DVNLPKADVD  ISGPKVDIEG
PDVNIEGPEG  KLKGPKLKMP  EMNIKAPKIS  MPDFDLHLKG  PKVKGDVDVS
LPKVEGDLKG  PEVDIKGPKV  DINAPDVGVQ  GPDWHLKMPK  VKMPKFSMPG
FKGEGPDGDV  KLPKADIDVS  GPKVDIEGPD  VNIEGPEGKL  KGPKFKMPEM
NIKAPKISMP  DIDLNLKGPK  VKGDVDVSLP  KVEGDLKGPE  VDIKGPKVDI
DAPDVDVHGP  DWHLKMPKIK  MPKISMPGFK  GEGPDVDVNL  PKADIDVSGP
KVDVECPDVN  IEGPEGKWKS  PKFKMPEMHF  KTPKISMPDI  DLNLTGPKIK
GDVDVTGPKV  EGDLKGPEVD  LKGPKVDIDV  PDVNVQGPDW  HLKMPKMKMP
KFSMPGFKAE  GPEVDVNLPK  ADVDVSGPKV  DVEGPDVNIE  GPEGKLKGPK
FKMPEMNIKA  PKIPMPDFDL  HLKGPKVKGD  VDISLPKVEG  DLKGPEVDIR
GPQVDIDVPD  VGVQGPDWHL  KMPKVKMPKF  SMPGFKGEGP  DVDVNLPKAD
LDVSGPKVDI  DVPDVNIEGP  EGKLKGPKFK  MPEMNIKAPK  ISMPDIDLNL
KGPKVKGDMD  VSLPKVEGDM  KVPDVDIKGP  KVDINAPDVD  VQGPDWHLKM
PKIKMPKISM  PGFKGEGPEV  DVNLPKADLD  VSGPKVDVDV  PDVNIEGPDA
KLKGPKFKMP  EMNIKAPKIS  MPDLDLNLKG  PKMKGEVDVS  LANVEGDLKG
PALDIKGPKI  DVDAPDIDIH  GPDAKLKGPK  LKMPDMHVNM  PKISMPEIDL
NLKGSKLKGD  VDVSGPKLEG  DIKAPSLDIK  GPEVDVSGPK  LNIEGKSKKS
RFKLPKFNFS  GSKVQTPEVD  VKGKKPDIDI  TGPKVDINAP  DVEVQGKVKG
SKFKMPFLSI  SSPKVSMPDV  ELNLKSPKVK  GDLDIAGPNL  EGDFKGPKVD
IKAPEVNLNA  PDVDVHGPDW  NLKMPKMKMP  KFSVSGLKAE  GPDVAVDLPK
GDINIEGPSM  NIEGPDLNVE  GPEGGLKGPK  FKMPDMNIKA  PKISMPDIDL
NLKGPKVKGD  VDISLPKLEG  DLKGPEVDIK  GPKVDINAPD  VDVHGPDWHL
KMPKVKMPKF  SMPGFKGEGP  EVDVTLPKAD  IDISGPNVDV  DVPDVNIEGP
DAKLKGPKFK  MPEMNIKAPK  ISMPDFDLNL  KGPKMKGDVV  VSLPKVEGDL
KGPEVDIKGP  KVDIDTPDIN  IEGSEGKFKG  PKFKIPEMHL  KAPKISMPDI
DLNLKGPKVK  GDVDVSLPKM  EGDLKGPEVD  IKGPKVDINA  PDVDVQGPDW
HLKMPKVKMP  KFSMPGFKGE  GPDVDVNLPK  ADLDVSGPKV  DIDVPDVNIE
GPEGKLKGPK  FKMPEMNIKA  PKISMPDIDL  NLKGPKVKGD  MDVSLPKVEG
DMQVPDLDIK  GPKVDINAPD  VDVRGPDWHL  KMPKIKMPKI  SMPGFKGEGP
EVDVNLPKAD  LDVSGPKVDV  DVPDVNIEGP  DAKLKGPKFK  MPEMNIKAPK
ISMPDFDLHL  KGPKVKGDVD  VSLPKMEGDL  KAPEVDIKGP  KVDIDAPDVD
VHGPDWHLKM  PKVKMPKFSM  PGFKGEGPEV  DVNLPKADID  VSGPKVDIDT
PDIDIHGPEG  KLKGPKFKMP  DLHLKAPKIS  MPEVDLNLKG  PKMKGDVDVS
LPKVEGDLKG  PEVDIKGPKV  DIDVPDVDVQ  GPDWHLKMPK  VKMPKFSMPG
FKGEGPDVDV  NLPKADLDVS  GPKVDIDVPD  VNIEGPDAKL  KGPKFKMPEM
NIKAPKISMP  DFDLHLKGPK  VKGDVDVSLP  KVEGDLKGPE  VDIKGPKVDI
DAPDVDVHGP  DWHLKMPKVK  MPKFSMPGFK  GEGPDVDVTL  PKADIEISGP
KVDIDAPDVS  IEGPDAKLKG  PKFKMPEMNI  KAPKISMPDI  DFNLKGPKVK
GDVDVSLPKV  EGDLKGPEID  IKGPSLDIDT  PDVNIEGPEG  KLKGPKFKMP
EMNIKAPKIS  MPDFDLHLKG  PKVKGDVDVS  LPKVESDLKG  PEVDIEGPEG
KLKGPKFKMP  DVHFKSPQIS  MSDIDLNLKG  PKIKGDMDIS  VPKLEGDLKG
PKVDVKGPKV  GIDTPDIDIH  GPEGKLKGPK  FKMPDLHLKA  PKISMPEVDL
NLKGPKVKGD  MDISLPKVEG  DLKGPEVDIR  DPKVDIDVPD  VDVQGPDWHL
KMPKVKMPKF  SMPGFKGEGP  DVDVNLPKAD  IDVSGPKVDV  DVPDVNIEGP
DAKLKGPKFK  MPEMSIKAPK  ISMPDIDLNL  KGPKVKGDVD  VTLPKVEGDL
KGPEADIKGP  KVDINTPDVD  VHGPDWHLKM  PKVKMPKFSM  PGFKGEGPDV
DVSLPKADID  VSGPKVDVDI  PDVNIEGPDA  KLKGPKFKMP  EINIKAPKIS
IPDVDLDLKG  PKVKGDFDVS  VPKVEGTLKG  PEVDLKGPRL  DFEGPDAKLS
GPSLKMPSLE  ISAPKVTAPD  VDLHLKAPKI  GFSGPKLEGG  EVDLKGPKVE
APSLDVHMDS  PDINIEGPDV  KIPKFKKPKF  GFGAKSPKAD  IKSPSLDVTV
PEAELNLETP  EISVGGKGKK  SKFKMPKIHM  SGPKIKAKKQ  GFDLNVPGGE
IDASLKAPDV  DVNIAGPDAA  LKVDVKSPKT  KKTMFGKMYF  PDVEFDIKSP
KFKAEAPLPS  PKLEGELQAP  DLELSLPAIH  VEGLDIKAKA  PKVKMPDVDI
SVPKIEGDLK  GPKVQANLGA  PDINIEGLDA  KVKTPSFGIS  APQVSIPDVN
VNLKGPKIKG  DVPSVGLEGP  DVDLQGPEAK  IKFPKFSMPK  IGIPGVKMEG
GGAEVHAQLP  SLEGDLRGPD  VKLEGPDVSL  KGPGVDLPSV  NLSMPKVSGP
DLDLNLKGPS  LKGDLDASVP  SMKVHAPGLN  LSGVGGKMQV  GGDGVKVPGI
DATTKLNVGA  PDVTLRGPSL  QGDLAVSGDI  KCPKVSVGAP  DLSLEASEGS
IKLPKMKLPQ  FGISTPGSDL  HVNAKGPQVS  GELKGPGVDV  NLKGPRISAP
NVDFNLEGPK  VKGSLGATGE  IKGPTVGGGL  PGIGVQGLEG  NLQMPGIKSS
GCDVNLPGVN  VKLPTGQISG  PEIKGGLKGS  EVGFHGAAPD  ISVKGPAFNM
ASPESDFGIN  LKGPKIKGGA  DVSGGVSAPD  ISLGEGHLSV  KGSGGEWKGP
QVSSALNLDT  SKFAGGLHFS  GPKVEGGVKG  GQIGLQAPGL  SVSGPQGHLE
SGSGKVTFPK  MKIPKFTFSG  RELVGREMGV  DVHFPKAEAS  IQAGAGDGEW
EESEVKLKKS  KIKMPKFNFS  KPKGKGGVTG  SPEASISGSK  GDLKSSKASL
GSLEGEAEAE  ASSPKGKFSL  FKSKKPRHRS  NSFSDEREFS  GPSTPTGTLE
FEGGEVSLEG  GKVKGKHGKL  KFGTFGGLGS  KSKGHYEVTG  SDDETGKLQG
SGVSLASKKS  RLSSSSSNDS  GNKVGIQLPE  VELSVSTKKE

2. Features

INTENDED USE
The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of AHNAK in human tissue homogenates, cell lysates and other biological fluids.

DETECTION RANGE
0.312-20ng/mL. The standard curve concentrations used for the ELISA’s were 20ng/mL, 10ng/mL, 5ng/mL, 2.5ng/mL, 1.25ng/mL, 0.625ng/mL, 0.312ng/mL.

SENSITIVITY
The minimum detectable dose of AHNAK is typically less than 0.117ng/mL.
The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.

SPECIFICITY
This assay has high sensitivity and excellent specificity for detection of AHNAK.
No significant cross-reactivity or interference between AHNAK and analogues was observed.
Note: Limited by current skills and knowledge, it is impossible to perform all possible cross-reactivity detection tests between AHNAK and all analogues, therefore, cross reactivity may still exist.

IMPORTANT NOTES
1. Limited by the current conditions and scientific technology, it is impossible to conduct comprehensive identification and analysis tests on the raw materials provided by suppliers. As a result, it is possible there are some qualitative and/or technical risks.
2. The final experimental results will be closely related to the validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available to obtain accurate results.
3. Kits from different batches may be a little different in detection range, sensitivity and color developing time. Please perform the experiment exactly according to the instruction manual included in your kit. Electronic ones on our website are for reference only.
4. Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
5. Protect all reagents from strong light during storage and incubation. All bottle caps of reagents should be closed tightly to prevent evaporation of liquids and contamination by microorganisms.
6. There may be a foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
7. Incorrect procedures during reagent preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
8. Even the same experimenter may get different results from two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before the general assay for each batch is recommended.
9. Each kit has undergone several rigorous quality control tests. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipment. Intra-assay variance among kits from different batches could arise from the above factors as well.
10. Kits from different manufacturers with the same item might produce different results, since we have not compared our products with other manufacturers.
11. The standard in this kit, as well as the antigens used in antibody preparation are typically recombinant proteins. Differently expressed sequences, expression systems, and/or purification methods can be used in the preparation of recombinant proteins. There is also the possibility of differences in the screening technique of antibodies and antibody pairs in our kits. As a result, we cannot guarantee that our kit will be able to detect recombinant proteins produced by other companies. We do NOT recommend using our ELISA kits for the detection of other recombinant proteins.
12. Validity period: 12 months.
13. The instruction manual also works with the 48T kit, but all reagents in the 48T kit are reduced by half.

You can reference link of the kit as following
https://dldevelop.com/Research-reagent/dl-ahnak-hu.html
https://www.dldevelop.com/uploadfile/data/DL-AHNAK-Hu.pdf

Introduction

Item	Standard		Test
Description	The kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of ROS1 in mouse serum, plasma, tissue homogenates or other biological fluids.		Conform
Identification	Colorimetric		Positive
Composition	Traditional ELISA Kit	Ready-to-Use ELISA KIT	Conform
	Pre-coated, ready to use 96-well strip plate 1	Pre-coated, ready to use 96-well strip plate 1
	Plate sealer for 96 wells 2	Plate sealer for 96 wells 2
	Standard 2	Standard 2
	Diluents buffer 1×45mL	Standard Diluent 1×20mL
	Detection Reagent A 1×120μL	Detection Solution A 1×12mL
	Detection Reagent B 1×120μL	Detection Solution B 1×12mL
	TMB Substrate 1×9mL	TMB Substrate 1×9mL
	Stop Solution 1×6mL	Stop Solution 1×6mL
	Wash Buffer (30 × concentrate) 1×20mL	Wash Buffer (30 × concentrate) 1×20mL
	Instruction manual 1	Instruction manual 1

Test principle

The microtiter plate provided in this kit has been pre-coated with an antibody specific to the index. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific to the index. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain the index, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of the index in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Recovery

Matrices listed below were spiked with certain level of recombinant ROS1 and the recovery rates were calculated by comparing the measured value to the expected amount of the index in samples.

Matrix	Recovery range (%)	Average(%)
serum(n=5)	81-93	86
EDTA plasma(n=5)	80-97	88
heparin plasma(n=5)	90-101	95

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of the index and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample	1：2	1：4	1：8	1：16
serum(n=5)	82-96%	83-98%	81-99%	93-101%
EDTA plasma(n=5)	88-101%	86-95%	90-102%	80-93%
heparin plasma(n=5)	80-91%	82-90%	95-104%	79-95%

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level the index were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level the index were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Stability

The stability of ELISA kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage conditions.
Note:
To minimize unnecessary influences on the performance, operation procedures and lab conditions, especially room temperature, air humidity and incubator temperatures should be strictly regulated. It is also strongly suggested that the whole assay is performed by the same experimenter from the beginning to the end.

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 2 hours at 37℃;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37℃;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 1 hour at 37℃;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37℃;
8. Add 50µL Stop Solution. Read at 450nm immediately.