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Human Deoxyribonuclease gamma (DNASE1L3) ELISA Kit (HUEB2365)

SKU:
HUEB2365
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q13609
Range:
0.312-20 ng/mL
ELISA Type:
Sandwich
Synonyms:
Deoxyribonuclease gamma, deoxyribonuclease I like 3, DHP2, DNAS1L3, DNase gamma, DNase I homolog protein DHP2, DNase I like 3, DNASE1L3, Liver and spleen DNase, LS DNase, LSD
Reactivity:
Human
€649
Frequently bought together:

Description

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Human Deoxyribonuclease gamma (DNASE1L3) ELISA Kit

The Human Deoxyribonuclease Gamma (DNase1L3) ELISA Kit is a powerful tool for accurately measuring levels of DNase1L3 in human biological samples such as serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit delivers precise and reproducible results, making it well-suited for a variety of research applications.DNase1L3, also known as deoxyribonuclease gamma, is a key enzyme involved in DNA degradation and clearance, playing a role in maintaining genomic integrity and regulating immune responses.

Dysregulation of DNase1L3 has been implicated in autoimmune diseases, inflammatory conditions, and certain cancers, highlighting its importance as a potent biomarker for studying these diseases and potential therapeutic targets.Overall, the Human DNase1L3 ELISA Kit offers researchers a reliable tool for investigating the role of DNase1L3 in various biological processes and disease states, advancing our understanding of its functions and therapeutic implications.

Product Name:Human Deoxyribonuclease gamma (DNASE1L3) ELISA Kit
SKU:HUEB2365
Size:96T
Target:Human Deoxyribonuclease gamma (DNASE1L3)
Synonyms:DNase I homolog protein DHP2, Deoxyribonuclease I-like 3, Liver and spleen DNase, DNase I-like 3, LS-DNase, DNase gamma, DHP2, DNAS1L3
Assay Type:Sandwich
Detection Method:ELISA
Reactivity:Human
Detection Range:0.312-20ng/mL
Sensitivity:0.18ng/mL
Intra CV:8.9%
Inter CV:11.5%
Linearity:
Sample1:21:41:81:16
Serum(N=5)93-102%95-106%93-103%106-115%
EDTA Plasma(N=5)100-110%100-110%93-103%83-96%
Heparin Plasma(N=5)80-91%102-112%99-111%107-118%
Recovery:
Sample TypeAverage(%)Recovery Range(%)
Serum9488-100
Plasma9690-102
Function:Has DNA hydrolytic activity. Is capable of both single- and double-stranded DNA cleavage, producing DNA fragments with 3'-OH ends (By similarity). Can cleave chromatin to nucleosomal units and cleaves nucleosomal and liposome-coated DNA (PubMed:9070308, PubMed:9714828, PubMed:14646506, PubMed:10807908, PubMed:27293190). Acts in internucleosomal DNA fragmentation (INDF) during apoptosis and necrosis (PubMed:23229555, PubMed:24312463). The role in apoptosis includes myogenic and neuronal differentiation, and BCR-mediated clonal deletion of self-reactive B cells (By similarity). Is active on chromatin in apoptotic cell-derived membrane-coated microparticles and thus suppresses anti-DNA autoimmunity (PubMed:27293190).
Uniprot:Q13609
Sample Type:Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
Specificity:Natural and recombinant human Deoxyribonuclease gamma
Research Area:Epigenetics
Subcellular Location:Nucleus Endoplasmic reticulum Secreted Translocates from the endoplasmic reticulum to the nucleus during apoptosis (PubMed:23229555). Contradictory reports exist about the subcellular localization under normal physiological conditions. Under conditions of cell death, may diffuse and/or be actively transported to the nucleus.
Storage:Please see kit components below for exact storage details
Note:For research use only
UniProt Protein Function:DNASE1L3: Has DNA hydrolytic activity. Does not bind to actin. Cleaves chromatin DNA to nucleosomal units. Defects in DNASE1L3 are the cause of systemic lupus erythematosus type 16 (SLEB16). A rare autosomal recessive form of systemic lupus erythematosus with childhood onset, characterized by high frequency of anti-neutrophil cytoplasmic antibodies and lupus nephritis. Systemic lupus erythematosus is a chronic, relapsing, inflammatory, and often febrile multisystemic disorder of connective tissue, characterized principally by involvement of the skin, joints, kidneys and serosal membranes. It is of unknown etiology, but is thought to represent a failure of the regulatory mechanisms of the autoimmune system. The disease is marked by a wide range of system dysfunctions, an elevated erythrocyte sedimentation rate, and the formation of LE cells in the blood or bone marrow. Belongs to the DNase I family.
UniProt Protein Details:

Protein type:EC 3.1.21.-; Secreted, signal peptide; Calcium-binding; Secreted; Deoxyribonuclease; Apoptosis

Chromosomal Location of Human Ortholog: 3p14.3

Cellular Component: nucleus

Molecular Function:endodeoxyribonuclease activity, producing 5'-phosphomonoesters; DNA binding; deoxyribonuclease activity; calcium ion binding; endodeoxyribonuclease activity

Biological Process: DNA fragmentation during apoptosis; DNA metabolic process

Disease: Systemic Lupus Erythematosus 16

NCBI Summary:This gene encodes a member of the deoxyribonuclease I family. The encoded protein hydrolyzes DNA, is not inhibited by actin, and mediates the breakdown of DNA during apoptosis. Mutations in this gene are a cause of systemic lupus erythematosus-16. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene. [provided by RefSeq, Feb 2012]
UniProt Code:Q13609
NCBI GenInfo Identifier:2494173
NCBI Gene ID:1776
NCBI Accession:Q13609.1
UniProt Related Accession:Q13609
Molecular Weight:33.4 kDa
NCBI Full Name:Deoxyribonuclease gamma
NCBI Synonym Full Names:deoxyribonuclease 1 like 3
NCBI Official Symbol:DNASE1L3
NCBI Official Synonym Symbols:LSD; DHP2; SLEB16; DNAS1L3
NCBI Protein Information:deoxyribonuclease gamma
UniProt Protein Name:Deoxyribonuclease gamma
UniProt Synonym Protein Names:DNase I homolog protein DHP2; Deoxyribonuclease I-like 3; DNase I-like 3; Liver and spleen DNase; LS-DNase; LSD
Protein Family:Deoxyribonuclease
UniProt Gene Name:DNASE1L3
UniProt Entry Name:DNSL3_HUMAN
Component Quantity (96 Assays) Storage
ELISA Microplate (Dismountable) 8×12 strips -20°C
Lyophilized Standard 2 -20°C
Sample Diluent 20ml -20°C
Assay Diluent A 10mL -20°C
Assay Diluent B 10mL -20°C
Detection Reagent A 120µL -20°C
Detection Reagent B 120µL -20°C
Wash Buffer 30mL 4°C
Substrate 10mL 4°C
Stop Solution 10mL 4°C
Plate Sealer 5 -

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

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

Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. 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 experiments. We recommend running all samples in duplicate.

Step
1. Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C.
2. Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform.
3. Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4. Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C.
5. Repeat the wash process for five times as conducted in step 3.
6. Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction.
7. Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
8. Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters.
9. After experiment, store all reagents according to the specified storage temperature respectively until their expiry.

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 Type Protocol
Serum If 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.
Plasma Collect 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 Fluid Collect 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 supernatant Collect 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 lysates Solubilize 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 homogenates The 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 lysates Rinse 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 Milk Collect 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.
ELISAAntibodies
Human DNASE1L3 (Deoxyribonuclease gamma) ELISA Kit (HUFI05304)Anti-DNASE1L3 Antibody (CAB15608)
DNASE1L1 Antibody (PACO08898)
DNAJB4 Antibody (PACO16195)
IKZF1 Antibody, Biotin conjugated (PACO35629)
DNASE1L3 Antibody (PACO35630)

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