Human Kallikrein 10 ELISA Kit

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Antigen
Kallikrein 10 (KLK10) ELISA Kits
  • KLK10
  • 2300002A13Rik
  • NES1
  • PRSSL1
  • kallikrein-related peptidase 10
  • kallikrein related-peptidase 10
  • KLK10
  • Klk10
Reactivity
Human
Alternatives
Kits with alternative reactivity to:
16
3
1
1
1
1
1
1
1
1
1
Method Type
Competition ELISA
Detection Range
617.3-50000 pg/mL
Minimum Detection Limit
617.3 pg/mL
Application
ELISA
Options
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Catalog No. ABIN415167
$ 530.53
Plus shipping costs $45.00
Relevance Score ABIN Method Type Sample Type Detection Range Detection Minimum Supplier References Details
9.864998 ABIN414758 Competition ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 123.46-10000 pg/mL 123.46 pg/mL Log in to see
9.864998 ABIN367338 Sandwich ELISA 1.675-40 ng/mL 1.675 ng/mL Log in to see
9.864998 ABIN2703281 Sandwich ELISA Plasma, Cell Culture Supernatant, Serum 2-600 ng/mL 2 ng/mL Log in to see
9.864998 ABIN4947807 Competition ELISA Plasma, Cell Lysate, Cell Culture Supernatant, Serum, Tissue Homogenate, Biological Fluids 117.2-30.000 pg/mL 117.2 pg/mL Log in to see
1 ABIN1029058 Sandwich ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 0.156-10 ng/mL 0.156 ng/mL Log in to see
1 ABIN848287 Sandwich ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate 23.44-1500 pg/mL 23.44 pg/mL Log in to see
1 ABIN455643 Sandwich ELISA Cell Culture Supernatant, Serum, Plasma, Tissue Homogenate, Biological Fluids 0.625-40 ng/mL 0.625 ng/mL Log in to see
1 ABIN626516 Competition ELISA Serum, Cell Culture Supernatant, Tissue Homogenate, Plasma, Body Fluids 50-1000 pg/mL 50 pg/mL Log in to see
1 ABIN5592286 Sandwich ELISA Biological Fluids, Plasma, Serum, Tissue Homogenate 0.625-40 ng/mL 0.625 ng/mL Log in to see
1 ABIN5526045 Sandwich ELISA Plasma, Serum, Biological Fluids, Tissue Homogenate 0.625-40 ng/mL 0.625 ng/mL Log in to see
1 ABIN2535745 Competition ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 123.46-10000 pg/mL 123.46 pg/mL Log in to see
1 ABIN4970601 Sandwich ELISA Cell Culture Supernatant, Plasma, Tissue Homogenate, Serum, Cell Lysate, Biological Fluids 0.156-10 ng/mL 0.156 ng/mL Log in to see
1 ABIN2535744 Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 35-2000 pg/mL 35 pg/mL Log in to see
1 ABIN5596627 Sandwich ELISA Log in to see
1 ABIN2948954 0.625-40 ng/mL 0.625 ng/mL Log in to see
1 ABIN2944382 117.19-30000 pg/mL 117.19 pg/mL Log in to see

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General

Antigen Kallikrein 10 (KLK10) ELISA Kits
Reactivity Human
Kits with alternative reactivity to:
(16), (3), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Method Type Competition ELISA
Detection Range 617.3-50000 pg/mL
Minimum Detection Limit 617.3 pg/mL
Application ELISA
Pubmed 14 references available
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Product Details Kallikrein 10 ELISA Kit

Target details Application Details Handling ProductDetails: References for Kallikrein 10 Kit (ABIN415167) Images
Purpose The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of NES1 in Serum,Plasma,Biological Fluids
Sample Type Biological Fluids, Plasma, Serum
Detection Method Colorimetric
Analytical Method Quantitative
Specificity

This assay has high sensitivity and excellent specificity for detection of Nesfatin 1 (NES1).
No significant cross-reactivity or interference between Nesfatin 1 (NES1) and analogues was observed.

Cross-Reactivity (Details) No significant cross-reactivity or interference between Nesfatin 1 (NES1) and analogues was observed.
Sensitivity 234.2 pg/mL
Components
  • Pre-coated, ready to use 96-well strip plate
  • Plate sealer for 96 wells
  • Standard Diluent
  • Assay Diluent A
  • Assay Diluent B
  • Stop Solution
  • Standard
  • Detection Reagent A
  • Detection Reagent B
  • TMB Substrate
  • Wash Buffer (30 × concentrate)
  • Instruction manual
Material not included
  • Microplate reader with 450 nm filter.
  • Precision single or multi-channel pipettes and disposable tips.
  • Eppendorf Tubes for diluting samples.
  • Deionized or distilled water.
  • Absorbent paper for blotting the microtiter plate.
  • Container for Wash Solution

Target details

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Antigen
Alternative Name NES1 (KLK10 ELISA Kit Abstract)
Pathways Complement System

Application Details

Product Details Kallikrein 10 ELISA Kit Target details Handling ProductDetails: References for Kallikrein 10 Kit (ABIN415167) Images back to top
Application Notes
  • Limited by the current condition and scientific technology, we cannot completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
  • The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
  • Kits from different batches may be a little different in detection range, sensitivity and color developing time.
  • Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
  • Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
  • There may be some 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.
  • Wrong operations during the reagents 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.
  • Even the same operator might get different results in 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 assay for each batch is recommended.
  • Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
  • Kits from different manufacturers for the same item might produce different results, since we have not compared our products with other manufacturers.
Comment

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Sample Volume 50 μL
Assay Time 2 h
Plate Pre-coated
Protocol This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Nesfatin 1 (NES1) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Nesfatin 1 (NES1) and unlabeled Nesfatin 1 (NES1) (Standards or samples) with the pre-coated antibody specific to Nesfatin 1 (NES1). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Nesfatin 1 (NES1) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Nesfatin 1 (NES1) in the sample.
Reagent Preparation
  • Bring all kit components and samples to room temperature (18-25°C) before use.
  • Standard - Reconstitute the Standard with 1.0mL of Standard Diluent, kept for 10 minutes at room temperature, shake gently(not to foam). The concentration of the standard in the stock solution is 150,000pg/mL. Please firstly dilute the stock solution to 50,000pg/mL and the diluted standard serves as the highest standard (50,000pg/mL). Then prepare 5 tubes containing 0.6mL Standard Diluent and produce a triple dilution series. Mix each tube thoroughly before the next transfer. Set up 5 points of diluted standard such as 50,000pg/mL, 16,666.7pg/mL, 5,555.6pg/mL, 1,851.9pg/mL, 617.3pg/mL, and the last EP tubes with Standard Diluent is the blank as 0pg/mL.
  • Assay Diluent A and Assay Diluent B - Dilute 6mL of Assay Diluent A or B Concentrate(2×) with 6mL of deionized or distilled water to prepare 12 mL of Assay Diluent A or B. (In fact, more than 6mL Assay Diluent A and Assay Diluent B are contained in the bottles. Therefore, in every test, please precisely pipette required amount of Diluent and make double dilution in a new container. The prepared working dilution cannot be frozen.)
  • Detection Reagent A and Detection Reagent B - Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute to the working concentration with working Assay Diluent A or B, respectively (1:100).
  • Wash Solution - Dilute 20mL of Wash Solution concentrate (30×) with 580mL of deionized or distilled water to prepare 600 mL of Wash Solution (1×).
  • TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.
Note:
  • Making serial dilution in the wells directly is not permitted.
  • Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37°C directly.
  • Detection Reagent A and B are sticky solutions, therefore, slowly pipette them to reduce the volume errors.
  • Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10µL for once pipetting.
  • The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
  • If crystals have formed in the Wash Solution concentrate (30×), warm to room temperature and mix gently until the crystals are completely dissolved.
  • Contaminated water or container for reagent preparation will influence the detection result.
Sample Collection Serum: Allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 20 minutes at approximately 1000 × g. Assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.

Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.

Biological Fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20 °C or -80 °C for later use. Avoid repeated freeze/thaw cycles.
Sample Preparation
  • We are only responsible for the kit itself, but not for the samples consumed during the assay. The user should calculate the possible amount of the samples used in the whole test. Please reserve sufficient samples in advance.
  • Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments. Sample should be diluted by 0.01mol/L PBS(PH=7.0-7.2).
  • If the samples are not indicated in the manual, a preliminary experiment to determine the validity of the kit is necessary.
  • Tissue or cell extraction samples prepared by chemical lysis buffer may cause unexpected ELISA results due to the impacts from certain chemicals.
  • Due to the possibility of mismatching between antigen from other origin and antibody used in our kits (e.g.antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.
  • Influenced by the factors including cell viability, cell number or sampling time, samples from cell culture supernatant may not be detected by the kit.
  • Fresh samples without long time storage is recommended for the test. Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.
Assay Procedure
  1. Prepare all reagents, samples and standards,
  2. Add 50μL standard or sample to each well.
    And then add 50μL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37 °C,
  3. Aspirate and wash 3 times,
  4. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
  5. Aspirate and wash 5 times,
  6. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
  7. Add 50μL Stop Solution. Read at 450 nm immediately.
Calculation of Results This assay employs the competitive inhibition enzyme immunoassay technique, so there is an inverse correlation between NES1 concentration in the sample and the assay signal intensity. Average the duplicate readings for each standard, control, and samples. Create a standard curve on log-log or semi-log graph paper, with the log of NES1 concentration on the y-axis and absorbance on the x-axis. Draw the best fit straight line through the standard points and it can be determined by regression analysis. Using some plot software, for instance, curve expert 1.30, is also recommended. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
In order to make the calculation easier, we plot the O.D. value of the standard (X-axis) against the log of concentration of the standard (Y-axis), although concentration is the independent variable and O.D. value is the dependent variable. The O.D. values of the standard curve may vary according to the conditions of assay performance (e.g. operator, pipetting technique, washing technique or temperature effects). Typical standard curve below is provided for reference only.
Assay Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Nesfatin 1 (NES1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Nesfatin 1 (NES1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Restrictions For Research Use only

Handling

Product Details Kallikrein 10 ELISA Kit Target details Application Details ProductDetails: References for Kallikrein 10 Kit (ABIN415167) Images back to top
Precaution of Use The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
Handling Advice

The stability of 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 condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Storage 4 °C
Storage Comment
  • For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
  • For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal.
    Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
  • For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.
Expiry Date 6 months

ProductDetails: References for Kallikrein 10 Kit (ABIN415167)

Product Details Kallikrein 10 ELISA Kit Target details Application Details Handling Images back to top
Product cited in:

Ustabaş Kahraman, Vehapoğlu, Özgen, Terzioğlu, Cesur, Dündaröz: "Correlation of Brain Neuropeptide (Nesfatin-1 and Orexin-A) Concentrations with Anthropometric and Biochemical Parameters in Malnourished Children." in: Journal of clinical research in pediatric endocrinology, Vol. 7, Issue 3, pp. 197-202, 2016

Cakir, Calikoglu, Yılmaz, Akpinar, Bayraktutan, Topcu: "Serum nesfatin-1 levels: a potential new biomarker in patients with subarachnoid hemorrhage." in: The International journal of neuroscience, pp. 1-7, 2016

Ahmadizad, Avansar, Ebrahim, Avandi, Ghasemikaram: "The effects of short-term high-intensity interval training vs. moderate-intensity continuous training on plasma levels of nesfatin-1 and inflammatory markers." in: Hormone molecular biology and clinical investigation, Vol. 21, Issue 3, pp. 165-73, 2015

Ülger, Gül, Uslukaya, O?uz, Bozda?, Yüksel, Böyük: "New hormones to predict the severity of gallstone-induced acute pancreatitis." in: The Turkish journal of gastroenterology : the official journal of Turkish Society of Gastroenterology, Vol. 25, Issue 6, pp. 714-7, 2015

Alp, Görmüş, Güdücü, Bozkurt: "Nesfatin-1 levels and metabolic markers in polycystic ovary syndrome." in: Gynecological endocrinology : the official journal of the International Society of Gynecological Endocrinology, Vol. 31, Issue 7, pp. 543-7, 2015

Araz, Yilmazel Ucar, Dorman, Bayraktutan, Yayla, Yilmaz, Acemoglu, Halici, Akgun: "Is There a Relationship between Obstructive Sleep Apnea Syndrome Severity and Nesfatin-1?" in: Respiration; international review of thoracic diseases, Vol. 90, Issue 2, pp. 105-10, 2015

Sahin, Sahin, Ural, Cure, Senturk, Tekin, Balik, Cure, Yuce, Kirbas: "Nesfatin-1 and Vitamin D levels may be associated with systolic and diastolic blood pressure values and hearth rate in polycystic ovary syndrome." in: Bosnian journal of basic medical sciences / Udruženje basičnih mediciniskih znanosti = Association of Basic Medical Sciences, Vol. 15, Issue 3, pp. 57-63, 2015

?engül, Dilbaz, Hal?c?, Ferah, Çad?rc?, Y?lmaz: "Decreased serum nesfatin-1 levels in endometriosis." in: European journal of obstetrics, gynecology, and reproductive biology, Vol. 177, pp. 34-7, 2014

Ademoglu, Gorar, Carl?oglu, Yaz?c?, Dellal, Berberoglu, Akdeniz, Uysal, Karakurt: "Plasma nesfatin-1 levels are increased in patients with polycystic ovary syndrome." in: Journal of endocrinological investigation, Vol. 37, Issue 8, pp. 715-9, 2014

Türko?lu, Böyük, Tanr?verdi, Gündüz, Dusak, Kaplan, Gümü?: "The potential role of BMI, plasma leptin, nesfatin-1 and ghrelin levels in the early detection of pancreatic necrosis and severe acute pancreatitis: a prospective cohort study." in: International journal of surgery (London, England), Vol. 12, Issue 12, pp. 1310-3, 2014

Aslan, Celik, Celik, Turkcuoglu, Yilmaz, Karaer, Simsek, Celik, Aydin: "Cord blood nesfatin-1 and apelin-36 levels in gestational diabetes mellitus." in: Endocrine, Vol. 41, Issue 3, pp. 424-9, 2012

Gunay, Tutuncu, Aydin, Dag, Abasli: "Decreased plasma nesfatin-1 levels in patients with generalized anxiety disorder." in: Psychoneuroendocrinology, Vol. 37, Issue 12, pp. 1949-53, 2012

Ari, Ozturk, Bez, Oktar, Erduran: "High plasma nesfatin-1 level in patients with major depressive disorder." in: Progress in neuro-psychopharmacology & biological psychiatry, Vol. 35, Issue 2, pp. 497-500, 2011

Johnson, Drugan, Miller, Evans: "38" in: , Vol. 1363, Issue Nucleic acids research, pp. 28-39, 1991

Images

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Supplier Images
ELISA image for Kallikrein 10 ELISA Kit (KLK10) (ABIN415167) Kallikrein 10 (KLK10) ELISA Kit