Lipocalin 2 ELISA 试剂盒
-
- 抗原 See all Lipocalin 2 (LCN2) ELISA试剂盒
- Lipocalin 2 (LCN2)
-
适用
- 人
- 检测方法
- Colorimetric
- 实验类型
- Sandwich ELISA
- 检测范围
- 79-5000 pg/mL
- 最低检测浓度
- 79 pg/mL
- 应用范围
- ELISA
- 原理
- The OmniKine? Human NGAL ELISA Kit contains the components necessary for quantitative determination of natural or recombinant Human NGAL concentrations within any experimental sample including cell lysates, serum and plasma. This particular immunoassay utilizes the quantitative technique of a "Sandwich" Enzyme-Linked Immunosorbent Assay (ELISA) where the target protein (antigen) is bound in a "sandwich" format by the primary capture antibodies coated to each well-bottom and the secondary detection antibodies added subsequently by the investigator. The capture antibodies coated to the bottom of each well are specific for a particular epitope on Human NGAL while the user-added detection antibodies bind to epitopes on the captured target protein. Amid each step of the procedure, a series of wash steps must be performed to ensure the elimination of non- specific binding between proteins to other proteins or to the solid phase. After incubation and "sandwiching" of the target antigen, a peroxidase enzyme is conjugated to the constant heavy chain of the secondary antibody (either covalently or via Avidin/Streptavidin-Biotin interactions), allowing for a colorimetric reaction to ensue upon substrate addition. When the substrate TMB (3, 3', 5, 5'-Tetramethylbenzidine) is added, the reaction catalyzed by peroxidase yields a blue color that is representative of the antigen concentration. Upon sufficient color development, the reaction can be terminated through addition of Stop Solution (2 N Sulfuric Acid) where the color of the solution will turn yellow. The absorbance of each well can then be read by a spectrophotometer, allowing for generation of a standard curve and subsequent determination of protein concentration.
- 品牌
- OmniKine™
- 样品类型
- Cell Lysate, Serum, Plasma
- Analytical Method
- Quantitative
- 特异性
- The Human NGAL ELISA Kit allows for the detection and quantification of endogenous levels of natural and/or recombinant Human NGAL proteins.
- 交叉反应 （详细）
- The Human NGAL ELISA is capable of recognizing both recombinant and naturally produced Human NGAL proteins. The antigens listed below were tested at 50 ng/mL and did not exhibit significant cross reactivity or interference. Human: Lipocalin-1, MMP-9 Murine: Lipocalin-2 Rat: Lipocalin-2
- 产品特性
- The Human NGAL ELISA Kit allows for the detection and quantification of endogenous levels of natural and/or recombinant Human NGAL proteins within the range of 79-5000 pg/mL.
- 组件
-
- Microstrips Coated w / Capture Antibody: 12 x 8-Well Microstrips
- Protein Standard: Lyophilized (100 ng), Red container
- Biotinylated Detection Antibody: Lyophilized, Yellow container
- 400x Streptavidin-HRP: 30 μL, Blue container
- Wash Buffer (10x): 50 mL, Clear containter
- Assay Diluent: 50 mL, Clear container
- Ready-to-Use Substrate: 12 mL, Brown container
- Stop Solution: 12 mL, Clear container
- Adhesive Plate Sealers: 4 Sheets
- Technical Manual 1 Manual
- 试剂未包括
-
The following materials and/or equipment are NOT provided in this kit but are necessary to successfully conduct the experiment:
Microplate reader able to measure absorbance at 450 nm (with correction wavelength set to 540 nm or 570 nm)
Micropipettes with capability of measuring volumes ranging from 1 μl to 1 mL
Deionized or sterile water
Squirt bottle, manifold dispenser, multichannel pipette reservoir or automated microplate washer
Graph paper or computer software capable of generating or displaying logarithmic functions
Absorbent paper or vacuum aspirator
Test tubes or microfuge tubes capable of storing ≥1 mL
Bench
top centrifuge (optional)
Bench
top vortex (optional)
Orbital shaker (optional) - Featured
- Discover our best selling LCN2 ELISA Kit
- Top Product
- Discover our top product LCN2 ELISA Kit
-
Lipocalin 2 (LCN2) ELISA Kit
LCN2 适用: 人 Colorimetric Sandwich ELISA 15.6-1000 pg/mL Plasma, Serum, Tissue Homogenate, Urine
Lipocalin 2 (LCN2) ELISA KitLCN2 适用: 人 Colorimetric Sandwich ELISA 0.15 ng/mL - 10 ng/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate, Urine
Lipocalin 2 (LCN2) ELISA KitLCN2 适用: 小鼠 Colorimetric Competition ELISA 12.5 pg/mL - 800 pg/mL Plasma, Serum, Tissue Homogenate, Urine
Lipocalin 2 (LCN2) ELISA KitLCN2 适用: 大鼠 Colorimetric Sandwich ELISA 0.78 ng/mL - 50 ng/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate, Urine
Lipocalin 2 (LCN2) ELISA KitLCN2 适用: Pig Colorimetric Sandwich ELISA 0.31 ng/mL - 20 ng/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate, Urine
Lipocalin 2 (LCN2) ELISA KitLCN2 适用: 人 AA 21-198 Colorimetric Sandwich ELISA 156-10000 pg/mL Cell Culture Supernatant, Plasma (heparin), Saliva, Serum, Urine
Lipocalin 2 (LCN2) ELISA KitLCN2 适用: 大鼠 AA 21-198 Colorimetric Sandwich ELISA 78-5000 pg/mL Cell Culture Supernatant, Plasma (heparin), Serum, Urine
Lipocalin 2 (LCN2) ELISA KitLCN2 适用: 小鼠 Colorimetric Sandwich ELISA 0.15 ng/mL - 10 ng/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate, Urine
Lipocalin 2 (LCN2) ELISA KitLCN2 适用: 犬 Colorimetric Sandwich ELISA 15.6 ng/mL - 1000 ng/mL Plasma, Serum, Urine
Lipocalin 2 (LCN2) ELISA KitLCN2 适用: 猴 Colorimetric Sandwich ELISA 0.313 ng/mL - 20 ng/mL Plasma, Serum, Tissue Homogenate
-
- 板类型
- Pre-coated
- 实验流程
- This particular immunoassay utilizes the quantitative technique of a Sandwich Enzyme-Linked Immunosorbent Assay (ELISA) where the target protein (antigen) is bound in a sandwich format by the primary capture antibodies coated to each well-bottom and the secondary detection antibodies added subsequently by the investigator. The capture antibodies coated to the bottom of each well are specific for a particular epitope on the Human NGAL cytokine while the user-added detection antibodies bind to epitopes on the captured target protein. Amid each step of the procedure, a series of wash steps must be performed to ensure the elimination of non-specific binding between proteins to other proteins or to the solid phase. After incubation and sandwiching of the target antigen, a peroxidase enzyme is conjugated to the constant heavy chain of the secondary antibody (either covalently or via Avidin/Streptavidin-Biotin interactions), allowing for a colorimetric reaction to ensue upon substrate addition. When the substrate TMB (3, 3’, 5, 5’- Tetramethylbenzidine) is added, the reaction catalyzed by peroxidase yields a blue color that is representative of the antigen concentration. Upon sufficient color development, the reaction can be terminated through addition of Stop Solution (2 N Sulfuric Acid) where the color of the solution will turn yellow. The absorbance of each well can then be read by a spectrophotometer, allowing for generation of a standard curve and subsequent determination of protein concentration.
- 样品制备
-
If samples are to be used within 24 hours, aliquot and store at 4 °C. If samples are to be used over a long period of time, aliquot and store between -20 °C and -80 °C, depending on the duration of storage.
Note: Samples containing a visible precipitate or pellet must be clarified prior to use in the assay.
Caution: Avoid repeated freeze/thaw cycles to prevent loss of biological activity of proteins in experimental samples.- Cell Lysate and Supernatants:
Remove large cell components via centrifugation and perform the assay. Cell lysates and supernatants require a dilution using Assay Diluent. A serial dilution may be performed to determine a suitable dilution factor for the sample. For future use of the sample, follow the sample storage guidelines stated above. - Serum:
Allow samples to clot in a serum separator tube (SST) for 30 minutes. After sufficient clotting, centrifuge at 1000 x g for 15 minutes and remove serum from SST in preparation for the assay. Serum samples require at least a 1:50 dilution using Assay Diluent. For future use of the sample, follow the storage guidelines above. - Plasma:
Use heparin, citrate or EDTA as an anticoagulant to gather plasma from original biological sample. After collection of the plasma, centrifuge for 15 minutes at 1000 x g. This step must be performed within 30 minutes of plasma collection. Plasma samples require at least a 1:50 dilution using Assay Diluent. Afterwards, perform the assay or for future use of the sample, follow the storage guidelines stated above.
- Cell Lysate and Supernatants:
- 实验流程
-
Note: If possible, all incubation steps should be performed on an orbital shaker to equilibrate solutions when added to the microplate wells. Also, all provided solutions should be at ambient temperature prior to use.
Note: Avoid adding solutions into wells at an angle, always keep pipette tip perpendicular to plate bottom.
Reconstitution of Provided Materials:1. Reconstitute the Biotin-Conjugated Detection Antibody in 67 µL of ddH₂O for a concentration of 180 µg/ml.
2. Reconstitute the Protein Standard in 100 µL of ddH₂O for a concentration of 340 ng/ml.
3. Dilute the 50 mL of 10x Wash Buffer in 450 mL of ddH2O for 500 mL of 1x Wash Buffer.
- 结果分析
-
Generation of Standard Curve and Interpretation of Data
1. Average the duplicate or triplicate readings for each standard, control and sample and subtract the average zero standard optical density.
2. Generate a standard curve by using Microsoft Excel or other computer software capable of establishing a 4- Parameter Logistic (4-PL) curve fit. If using Excel or an alternative graphing tool, plot the average optical density values in absorbance units (y-axis) against the known standard concentrations in pg/ml (x-axis). Note: Only use the values in which a noticeable gradient can be established. Afterwards, generate a best fit curve or trend-line through the plotted points via regression analysis. - 限制
- 仅限研究用
-
- 注意事项
-
Reagents provided in this kit may be harmful if ingested, inhaled or absorbed through the skin. Please carefully review the MSDS for each reagent before conducting the experiment.
Stop Solution contains 2 N Sulfuric Acid (H2SO4) and is an extremely corrosive agent. Please wear proper eye, hand and face protection when handling this material. When the experiment is finished, be sure to rinse the plate with copious amounts of running water to dilute the Stop Solution prior to disposing the plate. - 注意事项
-
This ELISA kit is intended for research purposes only, NOT diagnostic or clinical procedures of any kind.
Materials included in this kit should NOT be used past the expiration date on the kit label.
Reagents or substrates included in this kit should NOT be mixed or substituted with reagents or substrates from any other kits.
Variations in pipetting technique, washing technique, operator laboratory technique, kit age, incubation time or temperature may cause differences in binding affinity of the materials provided.
The assay is designed to eliminate interference and background by other cellular macromolecules or factors present within any biological samples. However, the possibility of background noise cannot be fully excluded until all factors have been tested using the assay kit.
Reagents provided in this kit may be harmful if ingested, inhaled or absorbed through the skin. Please carefully review the MSDS for each reagent before conducting the experiment.
Stop Solution contains 2 N Sulfuric Acid (H2SO4) and is an extremely corrosive agent. Please wear proper eye, hand and face protection when handling this material. When the experiment is finished, be sure to rinse the plate with copious amounts of running water to dilute the Stop Solution prior to disposing the plate. - 储存条件
- 4 °C
- 储存方法
-
Note: If used frequently, reagents may be stored at 4 °C.
- Unopened Kits: Store at 4 °C for 6 months.
- Microstrips Coated w/ Capture Antibody, 400x Streptavidin-HRP Wash Buffer (10x), Assay Diluent Ready-to-Use Substrate, Stop Solution: 6 Months at 4 °C
- Protein Standard, Biotinylated Detection Antibody: Lyophilized: 6 Months (if Reconstituted: 1 Month) at 4 °C
-
- 抗原 See all Lipocalin 2 (LCN2) ELISA试剂盒
- Lipocalin 2 (LCN2)
- 别名
- NGAL (LCN2 产品)
- 别名
- LCN2 ELISA Kit, 24p3 ELISA Kit, MSFI ELISA Kit, NGAL ELISA Kit, AW212229 ELISA Kit, Sip24 ELISA Kit, lipocalin 2 ELISA Kit, LCN2 ELISA Kit, Lcn2 ELISA Kit
- 背景
- Lipocalin-2/NGAL is an iron-trafficking protein involved in multiple processes such as apoptosis, innate immunity and renal development. The protein binds iron through association with 2, 5-dihydroxybenzoic acid (2, 5- DHBA), a siderophore that shares structural similarities with bacterial enterobactin, and delivers or removes iron from the cell, depending on the context. The iron-bound form (holo-24p3) is internalized following binding to the SLC22A17 (24p3R) receptor, leading to release of iron and subsequent increase of intracellular iron concentration. In contrast, association of the iron-free form (apo-24p3) with the SLC22A17 (24p3R) receptor is followed by association with an intracellular siderophore, iron chelation and iron transfer to the extracellular medium, thereby reducing intracellular iron concentration. Lipocalin-2/NGAL is involved in apoptosis due to interleukin- 3 (IL3) deprivation, the iron-loaded form increases intracellular iron concentration without promoting apoptosis, while the iron-free form decreases intracellular iron levels, inducing expression of the pro-apoptotic protein BCL2L11/BIM, resulting in apoptosis. This protein is also involved in innate immunity, possibly by sequestrating iron, leading to limit bacterial growth. As a homodimer, it is disulfide-linked. As a heterodimer, it is disulfide-linked with MMP9. Lipocalin-2/NGAL is expressed in bone marrow and in tissues that are prone to exposure to microorganisms. High expression is found in bone marrow as well as in uterus, prostate, salivary gland, stomach, appendix, colon, trachea and lung. It is not found in the small intestine or peripheral blood leukocytes. Expression is activated by the oncoprotein BCR-ABL, BCR-ABL misregulates expression via the JAK/STAT pathway and binding of STAT5A to the promoter. Source: Entrez Gene, Swiss-Prot
- 途径
- Cellular Response to Molecule of Bacterial Origin, Transition Metal Ion Homeostasis
-
