Bottom-up mass spectrometry-based protein evaluation strategies using protease digestion are routinely used to establish and characterize proteins with excessive specificity and sensitivity. Method efficiency is usually measured by sequence protection functionality and the entire variety of attribute peptides recognized, when in comparison with predicted databases. Limitations to generally used solvent-based digestion strategies at present employed embody lengthy digestion occasions (18-24 h or extra), resulting in protease autolysis, which additionally precludes automation, decreases sensitivity, and will increase each intra- and inter-day efficiency variability.
This report describes the event and validation of a easy, 5 min tryptic denaturing natural digestion (DOD) technique for use with tandem mass spectrometry in bottom-up protein identification and characterization. It has been evaluated throughout choose protein toxins and diagnostic scientific protein targets, considerably bettering digestion efficiency when in comparison with different solution-based and enzyme-immobilized strategies. The technique was in comparison with two at present used bottom-up strategies, the 24 h filter-aided pattern prep (FASP) and Flash Digest (1 and Four h) strategies.
Single proteins used to match the strategies included the ricin gentle chain, ricin heavy chain, ricin holotoxin, serotype A Clostridium botulinum toxin, Staphylococcus enterotoxin B, ribonuclease A, and thyroglobulin. In checks, throughout the proteins investigated, the 5 min DOD digestion technique resulted in sequence coverages starting from 55 to 100%, with comparatively excessive reproducibility and precision; outcomes have been higher than or equal to FASP technique outcomes and have been tremendously enhanced when in comparison with Flash technique outcomes.
Importantly, DOD technique intra- and inter-day precision was a lot improved as in comparison with outcomes for each FASP and Flash digestions. These knowledge indicated that the DOD technique, when in comparison with the FASP and Flash Digest strategies, dramatically diminished digestion time, whereas sustaining or bettering the flexibility to detect and characterize focused proteins, and diminished analytical variability for tryptic digestion, leading to markedly quicker and extra exact analyses.

Greater Pain Severity is Associated with Higher Glucocorticoid Levels in Hair Among a Cohort of People Living with HIV
Isotope sample deconvolution as a profitable various to calibration curve for software in wastewater-based epidemiology
An isotope sample deconvolution (IPD) quantification technique has been utilized for the willpower of 5 substances (amphetamine, benzoylecgonine, cocaine, methamphetamine and MDMA) in wastewater for the applying in wastewater-based epidemiology (WBE). A beforehand validated technique that used a calibration curve for quantification was modified to use IPD. The two approaches have been in contrast by way of analytical uncertainty in restoration research of high quality management samples, i.e. six wastewater samples from completely different geographical origins spiked at two focus ranges.
Both strategies have been dependable as they handed (z-score < 2) in an interlaboratory train. After 60 particular person determinations, IPD offered 11 outcomes exterior restoration limits (70-120%) whereas the earlier technique produced 31 antagonistic outcomes. All imply values for IPD have been correct whereas 6 out of 10 outcomes confirmed RSD values greater than 30% or recoveries exterior limits when utilizing the previous technique. Moreover, the calculated technique bias for the latter doubles that of IPD, which, in flip, makes the mixed uncertainty (u(c)) a lot greater.
ASAH2 Rabbit pAb |
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A12596-200ul | Abclonal | 200 ul | EUR 459 |
ASAH2 Rabbit pAb |
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A12596-20ul | Abclonal | 20 ul | EUR 183 |
ASAH2 Rabbit pAb |
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A12596-50ul | Abclonal | 50 ul | EUR 223 |
ASAH2 Rabbit pAb |
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A7985-100ul | Abclonal | 100 ul | EUR 308 |
ASAH2 Rabbit pAb |
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A7985-200ul | Abclonal | 200 ul | EUR 459 |
ASAH2 Rabbit pAb |
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A7985-20ul | Abclonal | 20 ul | EUR 183 |
ASAH2 Rabbit pAb |
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A7985-50ul | Abclonal | 50 ul | EUR 223 |
ASAH2 Antibody |
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24731-100ul | SAB | 100ul | EUR 390 |
ASAH2 Antibody |
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36240-100ul | SAB | 100ul | EUR 252 |
ASAH2 Antibody |
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1-CSB-PA076840 | Cusabio |
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Description: A polyclonal antibody against ASAH2. Recognizes ASAH2 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC;ELISA:1:1000-1:2000, WB:1:200-1:1000, IHC:1:25-1:100 |
ASAH2 Antibody |
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1-CSB-PA133826 | Cusabio |
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Description: A polyclonal antibody against ASAH2. Recognizes ASAH2 from Human, Mouse, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC;ELISA:1:1000-1:2000, WB:1:200-1:1000, IHC:1:25-1:100 |
ASAH2 Antibody |
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1-CSB-PA002170EA01HU | Cusabio |
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Description: A polyclonal antibody against ASAH2. Recognizes ASAH2 from Human, Rat. This antibody is Unconjugated. Tested in the following application: ELISA, WB, IHC, IF; Recommended dilution: WB:1:500-1:5000, IHC:1:100-1:500, IF:1:50-1:500 |
ASAH2 Antibody |
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3828-100 | Biovision | EUR 316 |
ASAH2 Antibody |
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3828-30T | Biovision | EUR 146 |
ASAH2 antibody |
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70R-12124 | Fitzgerald | 100 ug | EUR 403 |
Description: Rabbit polyclonal ASAH2 antibody |
ASAH2 Antibody |
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4743-002mg | ProSci | 0.02 mg | EUR 171.82 |
Description: ASAH2 Antibody: Sphingolipids are hydrolyzed by ceramidases to yield sphingosine and fatty acids. These ceramidases are classified according to the pH range that supports their optimal activity. ASAH2 is a neutral ceramidase and key regulator of sphingolipid signaling metabolites at the cell surface, catalyzing the hydrolysis of the N-acyl linkage of ceramide at an optimal pH of 6.5-8.5. ASAH2 is a type II integral membrane protein that can be cleaved to yield a soluble secreted protein and acts as a repressor of apoptosis both by reducing C16-ceramide, thereby preventing ceramide-induced apoptosis, and generating sphingosine. Sphingosine exerts both mitogenic and apoptosis-inducing activities, and its phosphorylated form functions as an intra- and intercellular second messenger. ASAH2 is ubiquitously expressed primarily expressed with higher levels in the intestine, kidney, skeletal muscle and heart. Recent studies indicate that ASAH2 encoded neutral ceramidase is a key enzyme for the catabolism of dietary sphingolipids and regulates the levels of bioactive sphingolipid metabolites in the intestinal tract. |
ASAH2 Antibody |
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4743-01mg | ProSci | 0.1 mg | EUR 436.42 |
Description: ASAH2 Antibody: Sphingolipids are hydrolyzed by ceramidases to yield sphingosine and fatty acids. These ceramidases are classified according to the pH range that supports their optimal activity. ASAH2 is a neutral ceramidase and key regulator of sphingolipid signaling metabolites at the cell surface, catalyzing the hydrolysis of the N-acyl linkage of ceramide at an optimal pH of 6.5-8.5. ASAH2 is a type II integral membrane protein that can be cleaved to yield a soluble secreted protein and acts as a repressor of apoptosis both by reducing C16-ceramide, thereby preventing ceramide-induced apoptosis, and generating sphingosine. Sphingosine exerts both mitogenic and apoptosis-inducing activities, and its phosphorylated form functions as an intra- and intercellular second messenger. ASAH2 is ubiquitously expressed primarily expressed with higher levels in the intestine, kidney, skeletal muscle and heart. Recent studies indicate that ASAH2 encoded neutral ceramidase is a key enzyme for the catabolism of dietary sphingolipids and regulates the levels of bioactive sphingolipid metabolites in the intestinal tract. |
ASAH2 Polyclonal Antibody, HRP Conjugated |
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A62107 | EpiGentek | 100 µg | EUR 570.55 |
Description: reagents widely cited |
ASAH2 Polyclonal Antibody, FITC Conjugated |
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A62108 | EpiGentek | 100 µg | EUR 570.55 |
Description: Ask the seller for details |
ASAH2 Polyclonal Antibody, Biotin Conjugated |
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A62109 | EpiGentek | 100 µg | EUR 570.55 |
Description: The best epigenetics products |
Anti-ASAH2 antibody |
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STJ110292 | St John's Laboratory | 100 µl | EUR 277 |
Anti-ASAH2 antibody |
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STJ114470 | St John's Laboratory | 100 µl | EUR 277 |
ASAH2 Conjugated Antibody |
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C36240 | SAB | 100ul | EUR 397 |
ASAH2 siRNA |
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20-abx900469 | Abbexa |
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ASAH2 siRNA |
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20-abx908269 | Abbexa |
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ASAH2 siRNA |
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20-abx908270 | Abbexa |
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ASAH2 Peptide |
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4743P | ProSci | 0.05 mg | EUR 164.75 |
Description: (CT) ASAH2 peptide |
ASAH2 Antibody, HRP conjugated |
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1-CSB-PA002170EB01HU | Cusabio |
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Description: A polyclonal antibody against ASAH2. Recognizes ASAH2 from Human. This antibody is HRP conjugated. Tested in the following application: ELISA |
ASAH2 Antibody, FITC conjugated |
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1-CSB-PA002170EC01HU | Cusabio |
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Description: A polyclonal antibody against ASAH2. Recognizes ASAH2 from Human. This antibody is FITC conjugated. Tested in the following application: ELISA |
ASAH2 Antibody, Biotin conjugated |
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1-CSB-PA002170ED01HU | Cusabio |
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Description: A polyclonal antibody against ASAH2. Recognizes ASAH2 from Human. This antibody is Biotin conjugated. Tested in the following application: ELISA |
ASAH2 cloning plasmid |
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CSB-CL865115HU-10ug | Cusabio | 10ug | EUR 474 |
Description: A cloning plasmid for the ASAH2 gene. |
ASAH2 Blocking Peptide |
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3828BP-50 | Biovision | EUR 153 |
ASAH2 Blocking Peptide |
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33R-10941 | Fitzgerald | 50 ug | EUR 191 |
Description: A synthetic peptide for use as a blocking control in assays to test for specificity of ASAH2 antibody, catalog no. 70R-12124 |
Mouse ASAH2 shRNA Plasmid |
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20-abx974501 | Abbexa |
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Rat ASAH2 shRNA Plasmid |
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20-abx987160 | Abbexa |
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Human ASAH2 shRNA Plasmid |
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20-abx961153 | Abbexa |
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ASAH2 Recombinant Protein (Mouse) |
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RP117410 | ABM | 100 ug | Ask for price |
ASAH2 Recombinant Protein (Rat) |
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RP191108 | ABM | 100 ug | Ask for price |
ASAH2 Recombinant Protein (Human) |
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RP036760 | ABM | 100 ug | Ask for price |
N-Acylsphingosine Amidohydrolase 2 (ASAH2) Antibody |
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20-abx303129 | Abbexa |
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N-Acylsphingosine Amidohydrolase 2 (ASAH2) Antibody |
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20-abx007107 | Abbexa |
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N-Acylsphingosine Amidohydrolase 2 (ASAH2) Antibody |
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abx025039-400ul | Abbexa | 400 ul | EUR 523 |
N-Acylsphingosine Amidohydrolase 2 (ASAH2) Antibody |
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abx025039-80l | Abbexa | 80 µl | EUR 286 |
N-Acylsphingosine Amidohydrolase 2 (ASAH2) Antibody |
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20-abx214010 | Abbexa |
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N-Acylsphingosine Amidohydrolase 2 (ASAH2) Antibody |
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20-abx214011 | Abbexa |
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N-Acylsphingosine Amidohydrolase 2 (ASAH2) Antibody |
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20-abx177710 | Abbexa |
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N-Acylsphingosine Amidohydrolase 2 (ASAH2) Antibody |
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20-abx173712 | Abbexa |
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N-Acylsphingosine Amidohydrolase 2 (ASAH2) Antibody (HRP) |
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20-abx303130 | Abbexa |
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N-Acylsphingosine Amidohydrolase 2 (ASAH2) Antibody (FITC) |
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20-abx303131 | Abbexa |
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N-Acylsphingosine Amidohydrolase 2 (ASAH2) Antibody (Biotin) |
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20-abx303132 | Abbexa |
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ASAH2 ELISA Kit (Human) (OKCD01831) |
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OKCD01831 | Aviva Systems Biology | 96 Wells | EUR 831 |
Description: Description of target: Hydrolyzes the sphingolipid ceramide into sphingosine and free fatty acid at an optimal pH of 6.5-8.5. Acts as a key regulator of sphingolipid signaling metabolites by generating sphingosine at the cell surface. Acts as a repressor of apoptosis both by reducing C16-ceramide, thereby preventing ceramide-induced apoptosis, and generating sphingosine, a precursor of the antiapoptotic factor sphingosine 1-phosphate. Probably involved in the digestion of dietary sphingolipids in intestine by acting as a key enzyme for the catabolism of dietary sphingolipids and regulating the levels of bioactive sphingolipid metabolites in the intestinal tract.;Species reactivity: Human;Application: ;Assay info: Assay Methodology: Quantitative Sandwich Immunoassay;Sensitivity: < 0.31 ng/mL |
ASAH2 ORF Vector (Human) (pORF) |
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ORF012254 | ABM | 1.0 ug DNA | EUR 354 |
Asah2 ORF Vector (Rat) (pORF) |
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ORF063704 | ABM | 1.0 ug DNA | EUR 506 |
Asah2 ORF Vector (Mouse) (pORF) |
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ORF039138 | ABM | 1.0 ug DNA | EUR 506 |
Asah2 sgRNA CRISPR Lentivector set (Mouse) |
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K3499601 | ABM | 3 x 1.0 ug | EUR 339 |
ASAH2 sgRNA CRISPR Lentivector set (Human) |
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K0130501 | ABM | 3 x 1.0 ug | EUR 339 |
Asah2 sgRNA CRISPR Lentivector set (Rat) |
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K7087001 | ABM | 3 x 1.0 ug | EUR 339 |
Human N-Acylsphingosine Amidohydrolase 2 (ASAH2) Protein |
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20-abx654471 | Abbexa |
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Asah2 sgRNA CRISPR Lentivector (Mouse) (Target 1) |
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K3499602 | ABM | 1.0 ug DNA | EUR 154 |
Asah2 sgRNA CRISPR Lentivector (Mouse) (Target 2) |
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K3499603 | ABM | 1.0 ug DNA | EUR 154 |
Asah2 sgRNA CRISPR Lentivector (Mouse) (Target 3) |
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K3499604 | ABM | 1.0 ug DNA | EUR 154 |
ASAH2 sgRNA CRISPR Lentivector (Human) (Target 1) |
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K0130502 | ABM | 1.0 ug DNA | EUR 154 |
ASAH2 sgRNA CRISPR Lentivector (Human) (Target 2) |
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K0130503 | ABM | 1.0 ug DNA | EUR 154 |
ASAH2 sgRNA CRISPR Lentivector (Human) (Target 3) |
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K0130504 | ABM | 1.0 ug DNA | EUR 154 |
Mouse ASAH2 (Neutral ceramidase) ELISA Kit (CUSTOM) |
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ELI-34287m | Lifescience Market | 96 Tests | EUR 865 |
Human ASAH2 (Neutral ceramidase) ELISA Kit (CUSTOM) |
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ELI-34330h | Lifescience Market | 96 Tests | EUR 824 |
Rat ASAH2 (Neutral ceramidase) ELISA Kit (CUSTOM) |
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ELI-48960r | Lifescience Market | 96 Tests | EUR 886 |
Asah2 sgRNA CRISPR Lentivector (Rat) (Target 1) |
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K7087002 | ABM | 1.0 ug DNA | EUR 154 |
Asah2 sgRNA CRISPR Lentivector (Rat) (Target 2) |
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K7087003 | ABM | 1.0 ug DNA | EUR 154 |
Asah2 sgRNA CRISPR Lentivector (Rat) (Target 3) |
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K7087004 | ABM | 1.0 ug DNA | EUR 154 |
ASAH2 3'UTR Luciferase Stable Cell Line |
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TU001220 | ABM | 1.0 ml | EUR 1521 |
Asah2 3'UTR GFP Stable Cell Line |
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TU250951 | ABM | 1.0 ml | Ask for price |
Asah2 3'UTR Luciferase Stable Cell Line |
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TU200951 | ABM | 1.0 ml | Ask for price |
Asah2 3'UTR GFP Stable Cell Line |
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TU152183 | ABM | 1.0 ml | Ask for price |
Asah2 3'UTR Luciferase Stable Cell Line |
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TU102183 | ABM | 1.0 ml | Ask for price |
ASAH2 3'UTR GFP Stable Cell Line |
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TU051220 | ABM | 1.0 ml | EUR 1521 |
ASAH2 Protein Vector (Human) (pPB-C-His) |
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PV049013 | ABM | 500 ng | EUR 481 |
ASAH2 Protein Vector (Human) (pPB-N-His) |
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PV049014 | ABM | 500 ng | EUR 481 |
ASAH2 Protein Vector (Human) (pPM-C-HA) |
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PV049015 | ABM | 500 ng | EUR 481 |
ASAH2 Protein Vector (Human) (pPM-C-His) |
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PV049016 | ABM | 500 ng | EUR 329 |
ASAH2 Protein Vector (Human) (pPB-His-MBP) |
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PV324138 | ABM | 500 ng | EUR 481 |
ASAH2 Protein Vector (Human) (pPB-His-GST) |
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PV324139 | ABM | 500 ng | EUR 481 |
ASAH2 Protein Vector (Rat) (pPB-C-His) |
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PV254814 | ABM | 500 ng | EUR 1166 |
ASAH2 Protein Vector (Rat) (pPB-N-His) |
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PV254815 | ABM | 500 ng | EUR 1166 |
ASAH2 Protein Vector (Rat) (pPM-C-HA) |
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PV254816 | ABM | 500 ng | EUR 1166 |
ASAH2 Protein Vector (Rat) (pPM-C-His) |
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PV254817 | ABM | 500 ng | EUR 1166 |
ASAH2 Protein Vector (Mouse) (pPB-C-His) |
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PV156550 | ABM | 500 ng | EUR 1065 |
ASAH2 Protein Vector (Mouse) (pPB-N-His) |
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PV156551 | ABM | 500 ng | EUR 1065 |
ASAH2 Protein Vector (Mouse) (pPM-C-HA) |
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PV156552 | ABM | 500 ng | EUR 1065 |
ASAH2 Protein Vector (Mouse) (pPM-C-His) |
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PV156553 | ABM | 500 ng | EUR 1065 |
Human N-Acylsphingosine Amidohydrolase 2 (ASAH2) ELISA Kit |
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20-abx156834 | Abbexa |
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Human N-Acylsphingosine Amidohydrolase 2 (ASAH2) CLIA Kit |
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20-abx494694 | Abbexa |
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Human N-Acylsphingosine Amidohydrolase 2 (ASAH2) ELISA Kit |
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SEE689Hu-10x96wellstestplate | Cloud-Clone | 10x96-wells test plate | EUR 4731.5 |
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human N-Acylsphingosine Amidohydrolase 2 (ASAH2) in serum, plasma, tissue homogenates and other biological fluids. |
Human N-Acylsphingosine Amidohydrolase 2 (ASAH2) ELISA Kit |
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SEE689Hu-1x48wellstestplate | Cloud-Clone | 1x48-wells test plate | EUR 477.3 |
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human N-Acylsphingosine Amidohydrolase 2 (ASAH2) in serum, plasma, tissue homogenates and other biological fluids. |
Human N-Acylsphingosine Amidohydrolase 2 (ASAH2) ELISA Kit |
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SEE689Hu-1x96wellstestplate | Cloud-Clone | 1x96-wells test plate | EUR 639 |
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human N-Acylsphingosine Amidohydrolase 2 (ASAH2) in serum, plasma, tissue homogenates and other biological fluids. |
Human N-Acylsphingosine Amidohydrolase 2 (ASAH2) ELISA Kit |
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SEE689Hu-5x96wellstestplate | Cloud-Clone | 5x96-wells test plate | EUR 2575.5 |
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Human N-Acylsphingosine Amidohydrolase 2 (ASAH2) in serum, plasma, tissue homogenates and other biological fluids. |
Consequently, a easy change of information treatment-IPD quantification methodology-resulted in a decrease uncertainty of the estimated illicit drug focus, one of many principal steps contributing to the ultimate uncertainty within the normalized each day drug consumption by means of WBE. The present research demonstrated that the employment of IPD may also be very attention-grabbing for future purposes of WBE, particularly when matrix results are excessive, complicating correct quantification. In addition, when a excessive variety of samples and/or compounds have to be analysed, IPD is quicker than calibration and, finally, cost-effective when isotopically labelled inner customary is very costly.