Human IL-1 Antibody Bead Kit
Contents
1. invitrogen PRAS40 pT246 Singleplex Bead Kit Technical Data Sheet Catalog LHO0421 Lot 1013068 Note A letter at the end of the lot number signifies an additional packaging of this same lot Intended Use This kit is comprised of components for the measurement of PRAS40 pT246 in cell lysates and tissue homogenates Buffer reagents needed to complete the reaction are sold separately under Catalog LHB0002 These reagents are intended for use in the Luminex 100 200 and the FlexMAP 3D System This kit is configured for research use only and is not to be used in diagnostic procedures Note This kit uses a Co Incubation assay format Refer to the User Manual included in the Intracellular Protein Buffer Reagent Kit Cat LHB0002 for procedure This Singleplex Bead Kit has been validated for multiplexing with the following Singleplex Bead Kits available from Invitrogen Akt pS473 Cat LHOO101 p70S6K pTpS42 1 424 Cat LHO0191 GSK 38 pS9 Cat LHO0461 ERK1 2 pTpY185 187 Cat LHO0241 IRS 1 pS312 Cat LHO0521 ERK1 2 pT pY185 187 requires Sample Treatment Refer to the User Manual included in the Intracellular Protein Buffer Reagent Kit Cat LHB0002 for procedure Reagents Provided PRAS40 pT246 i Part LM118 Description Antibody Bead Concentrate 10X Lot 1022983 Size 0 25 mL 100 tests Bead Region 74 Form 0 25 mL 10X bead concentrate solution in storage buffer Contains 0 05 sodium az2. Carlsbad California 92008 PRAS40 pT246 Singleplex Bead Kit Technical Data Sheet Performance Characteristics Typical Standard Curve PRAS40 pT246 Standard Curve PRAS40 pT246 Sensitivity units mL 0 191 7500 6000 Recovery 10 FNN0021 98 Intra assay variation n 14 6 3 4500 MFI 3000 Inter assay variation n 32 7 4 Analytical Sensitivity The analytical sensitivity was determined by adding two standard deviations to the median fluorescence intensity obtained when the zero standard was assayed 32 times 1500 ss Recovery To evaluate recovery analytes were spiked at 3 different concentrations 0 100 200 300 400 into 10 Cell Extraction Buffer Invitrogen Cat FNN0011 and 10 NP40 Cell unnai Lysis Buffer Invitrogen Cat FNN0021 Note This is a representative standard curve Absolute MFI values will vary Species Reactivity Each analyte specific component of this reagent set recognizes human mouse and rat proteins Other species have not been tested Sample Preparation This kit has been validated with cell lysates prepared in Cell Extraction Buffer Invitrogen Cat FNN0011 and NP40 Cell Lysis Buffer Invitrogen Cat FNN0021 Optimization of cell stimulation and cell lysis procedures may be required for each specific application Refer to Product Insert for detailed instructions Important Note With some of the Singleplex bead kits available from Invitrogen cell
3. lysates must be pre incubated in Sample Treatment Buffer Invitrogen Cat STOO1 to optimize signal This sample pre incubation step has been found to adversely impact the signal obtained with other kits The impact of the Sample Treatment Buffer pre incubation step must therefore be considered when developing multiplexed assays for the detection of multiple markers with these reagents Effect of Sample Treatment Buffer on PRAS40 PRAS40 pT246 expression in non pT246 signal in cell lysates 200 pg mL treated and insulin treated MCF7 cells 15000 _10000 5000 0 Non treated Insulin Example Data To evaluate the performance of this kit MCF7 cells The data presented above demonstrate the impact of the Sample were serum starved for 15 hours and either left untreated or treated Treatment Buffer pre incubation step on the observed signal In this with 100 nM insulin for 15 minutes The results indicated that study PMA stimulated Jurkat cells were lysed in Cell Extraction Buffer phosphorylation of PRAS40 at threonine 246 increased following Lysates were either treated with Sample Treatment Buffer STB or the insulin treatment Sample Treatment Buffer incubation step was omitted STB Explanation of symbols Catalogue Number Batch code RUS Research Use Only IVD In vitro diagnostic medical device ti tenutecturer EC REP European Community authorised representative H Without does not contain Wit
4. e lot specific Please verify all concentration values entered in data analysis software Reconstitution Reconstitute in 1 0 mL Assay Diluent Recommended Starting Concentration for Standard Curve Upon reconstitution the starting concentration of standard is the value cited above Make serial 1 2 dilutions in Assay Diluent Refer to the User Manual included in the buffer kit for further information For Research Use Only CAUTION Not for human or animal therapeutic or diagnostic use Invitrogen Corporation 7335 Executive Way Frederick MD 21704 Tel 800 955 6288 E mail techsupport invitrogen com LHO0421 Page 1 of 2 MAN0004968 Rev 1 00 Effective Date 24 AUG 2011 Limited Use Label License Research Use Only The purchase of this product conveys to the purchaser the limited non transferable right to use the purchased amount of the product only to perform internal research for the sole benefit of the purchaser No right to resell this product or any of its components is conveyed expressly by implication or by estoppel This product is for internal research purposes only and is not for use in commercial applications of any kind including without limitation quality control and commercial services such as reporting the results of purchaser s activities for a fee or other form of consideration For information on obtaining additional rights please contact outlicensing lifetech com or Out Licensing Life Technologies 5791 Van Allen Way
5. h contains rote Q Protect from light Consult accompanying documents m Ligi A Directs the user to consult instructions for use IFU accompanying the product For Research Use Only CAUTION Not for human or animal therapeutic or diagnostic use www invitrogen com Invitrogen Corporation 7335 Executive Way Frederick MD 21704 Tel 800 955 6288 E mail techsupport invitrogen com LHO0421 Page 2 of 2 MAN0004968 Rev 1 00 Effective Date 24 AUG 2011 Limited Use Label License Research Use Only The purchase of this product conveys to the purchaser the limited non transferable right to use the purchased amount of the product only to perform internal research for the sole benefit of the purchaser No right to resell this product or any of its components is conveyed expressly by implication or by estoppel This product is for internal research purposes only and is not for use in commercial applications of any kind including without limitation quality control and commercial services such as reporting the results of purchaser s activities for a fee or other form of consideration For information on obtaining additional rights please contact outlicensing lifetech com or Out Licensing Life Technologies 5791 Van Allen Way Carlsbad California 92008
6. ide as a preservative Storage Light sensitive material Store at 2 to 8 C in the dark until the expiration date indicated on the kit PRAS40 pT246 2 Part DNI18 Description Detection Ab Concentrate 10X Lot 1022981 Size 0 5 mL 100 tests Form 0 5 mL of a 10X stock of Detector Antibody Concentrate Contains 0 1 sodium azide as preservative Concentration of antibody is matched to this lot of beads Do not mix lots of Coated Beads and Detector Antibody Storage Store at 2 to 8 C until the expiration date indicated on the kit 3 Part SM040 Description Akt Pathway Phospho 7 Plex Standard Lot 1007684 Size 2 Vials Form Lyophilized The proteins in this standard have been calibrated against the masses of highly purified recombinant proteins with the respective Invitrogen ELISA kits when available Contains 0 1 sodium azide as a preservative Storage Store at 2 to 8 C Use within 1 hour after reconstitution The remaining reconstituted standard should be discarded or frozen in aliquots at 80 C for further use The standard can be frozen and thawed one time without loss of immunoreactivity Concentration of Reconstituted Standard IR pYpY 1162 1163 1609 units mL PRAS40 pT246 997 units mL IGF 1R pYpY1135 1136 1274 units mL p70S6K pTpS421 424 518 units mL IRS 1 pS312 503 units mL GSK 36 pS9 68 units mL Akt pS473 105 units mL Important note The concentrations of reconstituted standard ar
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