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Human ULK1(Unc-51 Like Kinase 1) ELISA Kit (HUFI03341)

SKU:
HUFI03341
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
O75385
Sensitivity:
9.375pg/ml
Range:
15.625-1000pg/ml
ELISA Type:
Sandwich
Synonyms:
ATG1, KIAA0722, ULK1, Unc 51 like kinase 1, UNC51, Unc51.1
Reactivity:
Human
Research Area:
Autophagy
€599
Frequently bought together:

Description

Human ULK1 (Unc-51 Like Kinase 1) ELISA Kit

The Human ULK1 (UNC-51-like kinase 1) ELISA Kit is specifically designed for the precise quantification of ULK1 levels in human serum, plasma, and cell culture supernatants. This kit boasts exceptional sensitivity and specificity, guaranteeing accurate and consistent results, which makes it an invaluable tool for a diverse array of research endeavors.ULK1, also known as UNC-51-like kinase 1, is a critical protein involved in autophagy, a cellular process essential for maintaining cellular homeostasis and removing damaged or unnecessary components.

Dysregulation of autophagy and ULK1 has been implicated in various diseases, including cancer, neurodegenerative disorders, and metabolic conditions. Therefore, ULK1 serves as a pivotal biomarker for investigating these diseases and exploring potential therapeutic interventions.Overall, the Human ULK1 ELISA Kit provides researchers with a robust and reliable means to study ULK1 levels, paving the way for a deeper understanding of disease mechanisms and the development of novel treatment strategies.

Product Name:Human ULK1(Unc-51 Like Kinase 1) ELISA Kit
Product Code:HUFI03341
Size:96 Assays
Alias:ATG1, KIAA0722, ULK1, Unc 51 like kinase 1, UNC51, Unc51.1
Detection method:Sandwich ELISA, Double Antibody
Application:This immunoassay kit allows for the in vitro quantitative determination of Human ULK1 concentrations in serum plasma and other biological fluids.
Sensitivity:9.375pg/ml
Range:15.625-1000pg/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Human ULK1 and the recovery rates were calculated by comparing the measured value to the expected amount of Human ULK1 in samples. Enquire for more information.
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human ULK1 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Enquire for more information.
CV(%):Intra-Assay: CV<8%
Inter-Assay: CV<10%
ComponentQuantityStorage
ELISA Microplate (Dismountable)8×12 strips4°C for 6 months
Lyophilized Standard24°C/-20°C
Sample/Standard Dilution Buffer20ml4°C
Biotin-labeled Antibody(Concentrated)120ul4°C (Protect from light)
Antibody Dilution Buffer10ml4°C
HRP-Streptavidin Conjugate(SABC)120ul4°C (Protect from light)
SABC Dilution Buffer10ml4°C
TMB Substrate10ml4°C (Protect from light)
Stop Solution10ml4°C
Wash Buffer(25X)30ml4°C
Plate Sealer5-

Other materials and equipment required:

  • Microplate reader with 450 nm wavelength filter
  • Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
  • Incubator
  • Deionized or distilled water
  • Absorbent paper
  • Buffer resevoir
UniprotO75385
UniProt Protein Function:ULK1: Serine/threonine-protein kinase involved in autophagy in response to starvation. Acts upstream of phosphatidylinositol 3- kinase PIK3C3 to regulate the formation of autophagophores, the precursors of autophagosomes. Part of regulatory feedback loops in autophagy: acts both as a downstream effector and negative regulator of mammalian target of rapamycin complex 1 (mTORC1) via interaction with RPTOR. Activated via phosphorylation by AMPK and also acts as a regulator of AMPK by mediating phosphorylation of AMPK subunits PRKAA1, PRKAB2 and PRKAG1, leading to negatively regulate AMPK activity. May phosphorylate ATG13/KIAA0652 and RPTOR; however such data need additional evidences. Plays a role early in neuronal differentiation and is required for granule cell axon formation. Interacts with GABARAP and GABARAPL2. Interacts (via C- terminus) with ATG13/KIAA0652. Part of a complex consisting of ATG13/KIAA0652, ULK1 and RB1CC1. Associates with the mammalian target of rapamycin complex 1 (mTORC1) through an interaction with RPTOR; the association depends on nutrient conditions and is reduced during starvation. Ubiquitously expressed. Detected in the following adult tissues: skeletal muscle, heart, pancreas, brain, placenta, liver, kidney, and lung. Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. APG1/unc-51/ULK1 subfamily.
UniProt Protein Details:Protein type:Protein kinase, Other; Autophagy; Kinase, protein; Protein kinase, Ser/Thr (non-receptor); EC 2.7.11.1; Other group; ULK family

Chromosomal Location of Human Ortholog: 12q24.3

Cellular Component: autophagic vacuole; cytosol; pre-autophagosomal structure membrane

Molecular Function:protein binding; protein complex binding; protein serine/threonine kinase activity; Rab GTPase binding

Biological Process: axon extension; cellular response to nutrient levels; macroautophagy; negative regulation of protein complex assembly; neurite development; peptidyl-serine phosphorylation; positive regulation of autophagy; positive regulation of macroautophagy; protein amino acid autophosphorylation; protein localization; regulation of autophagy; response to starvation

UniProt Code:O75385
NCBI GenInfo Identifier:317373288
NCBI Gene ID:8408
NCBI Accession:O75385.2
UniProt Secondary Accession:O75385,Q9UQ28,
UniProt Related Accession:O75385
Molecular Weight:112,631 Da
NCBI Full Name:Serine/threonine-protein kinase ULK1
NCBI Synonym Full Names:unc-51 like autophagy activating kinase 1
NCBI Official Symbol:ULK1  
NCBI Official Synonym Symbols:ATG1; ATG1A; UNC51; hATG1; Unc51.1  
NCBI Protein Information:serine/threonine-protein kinase ULK1
UniProt Protein Name:Serine/threonine-protein kinase ULK1
UniProt Synonym Protein Names:Autophagy-related protein 1 homolog; ATG1; hATG1; Unc-51-like kinase 1
Protein Family:Serine/threonine-protein kinase
UniProt Gene Name:ULK1  
UniProt Entry Name:ULK1_HUMAN

*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.

Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

StepProtocol
1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4.Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37 °C for 90 min.
6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample TypeProtocol
SerumIf using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

PlasmaCollect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal FluidCollect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell culture supernatantCollect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell lysatesSolubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Tissue homogenatesThe preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C.
Tissue lysatesRinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast MilkCollect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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