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Genome-wide DNA methylation profiling using Infinium assay

Genome-wide DNA methylation profiling utilizing Infinium® assay.

 

OBJECTIVE
Bisulfite sequence evaluation of particular person CpG websites inside genomic DNA is a robust method for methylation evaluation within the genome.
The main limitation of bisulfite-based strategies is parallelization. Each array and next-generation sequencing know-how are succesful of addressing this bottleneck. On this report, we describe the applying of Infinium® genotyping know-how to investigate bisulfite-converted DNA to concurrently question the methylation state of over 27,000 CpG websites from promoters of consensus coding sequences (CCDS) genes.
METHODS
We tailored the Infinium genotyping assay to readout an array of over 27,000 pairs of CpG methylation-specific question probes complementary to bisulfite-converted DNA. Two probes have been designed to every CpG website: a ‘methylated’ and an ‘unmethylated’ question probe.
The probe design assumed that every one underlying CpG websites have been ‘in section’ with the queried CpG website as a consequence of their shut proximity. Bisulfite conversion was carried out with a modified model of the Zymo EZ DNA Methylation™ package.
RESULTS
We utilized this know-how to measuring methylation ranges throughout a panel of 14 totally different human tissues, 4 Coriell cell strains and 6 most cancers cell strains.
We noticed that CpG websites inside CpG islands (CGIs) have been largely unmethylated throughout all tissues (~80% websites unmethylated, β < 0.2), whereas CpG websites in non-CGIs have been reasonably to extremely methylated (solely ~12% websites unmethylated, β < 0.2). Inside CGIs, solely roughly 3-6% of the loci have been extremely methylated; in distinction, exterior of CGIs roughly 25-40% of loci have been extremely methylated. Furthermore, tissue-specific methylation (variation in methylation throughout tissues) was far more prevalent in non-CGIs than inside CGIs.
CONCLUSIONS
Our outcomes display a genome-wide scalable array-based methylation readout platform that’s each extremely reproducible and quantitative. Within the close to future, this platform ought to allow the evaluation of lots of of 1000’s to tens of millions of CpG websites per pattern.
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I094 1000 ml
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AR1016 100mL (for 10000-15000 assays)
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abx098961-6ml 6 ml
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abx098962-12ml 12 ml
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1Z0216-100ml 100 ml
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1Z0216-1L 1 Liter
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42R-TB105 1 liter
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Sample diluent buffer

AR1106-1 30ml
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AR1106-2 12ml
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AR1106-3 12ml
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abx098951-20ml 20 ml
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abx296001-20ml 20 ml
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EpiQuik General Protein-DNA Binding Assay Kit (Colorimetric)

P-2004 96 Assays
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Description: kits suitable for this type of research

EpiQuik General Protein-DNA Binding Assay Kit (Fluorometric)

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85R-108 250 ml
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Description: Ready-to-use biotin free reagent applicable for blocking ELISA plates

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35-RC15 100 ml
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Description: Rheumatoid Factor Control Diluent

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SU11248 is a novel FLT3 tyrosine kinase inhibitor with potent exercise in vitro and in vivo.

FLT3 (fms-related tyrosine kinase/Flk2/Stk-2) is a receptor tyrosine kinase (RTK) primarily expressed on hematopoietic cells. In blasts from acute myelogenous leukemia (AML) sufferers, 2 lessons of FLT3 activating mutations have been recognized: inside tandem duplication (ITD) mutations within the juxtamembrane area (25%-30% of sufferers) and level mutations within the kinase area activation loop (7%-8% of sufferers).
FLT3-ITD mutations are probably the most frequent molecular defect recognized in AML and have been proven to be an unbiased prognostic issue for decreased survival. FLT3-ITD is due to this fact a beautiful molecular goal for remedy.
SU11248 is a lately described selective inhibitor with selectivity for cut up kinase area RTKs, together with platelet-derived development issue receptors, vascular endothelial development issue receptors, and KIT.
We present that SU11248 additionally has potent exercise in opposition to wild-type FLT3 (FLT3-WT), FLT3-ITD, and FLT3 activation loop (FLT3-Asp835) mutants in phosphorylation assays. SU11248 inhibits FLT3-driven phosphorylation and induces apoptosis in vitro. As well as, SU11248 inhibits FLT3-induced VEGF manufacturing. The in vivo efficacy of SU11248 was investigated in 2 FLT3-ITD fashions: a subcutaneous tumor xenograft mannequin and a bone marrow engraftment mannequin. We present that SU11248 (20 mg/kg/d) dramatically regresses FLT3-ITD tumors within the subcutaneous tumor xenograft mannequin and prolongs survival within the bone marrow engraftment mannequin.
Pharmacokinetic and pharmacodynamic evaluation in subcutaneous tumors confirmed {that a} single administration of an efficacious drug dose potently inhibits FLT3-ITD phosphorylation for as much as 16 hours following a single dose. These outcomes counsel that additional exploration of SU11248 exercise in AML sufferers is warranted.

Age-associated traits of murine hematopoietic stem cells.

Little is understood of age-associated useful modifications in hematopoietic stem cells (HSCs). We studied growing old HSCs on the clonal stage by isolating CD34(-/low)c-Equipment(+)Sca-1(+) lineage marker-negative (CD34(-)KSL) cells from the bone marrow of C57BL/6 mice. A inhabitants of CD34(-)KSL cells progressively expanded as age elevated.
No matter age, these cells shaped in vitro colonies with stem cell issue and interleukin (IL)-Three however not with IL-Three alone. They didn’t kind day 12 colony-forming unit (CFU)-S, indicating that they’re primitive cells with myeloid differentiation potential.
An in vivo limiting dilution assay revealed that numbers of multilineage repopulating cells elevated twofold from 2 to 18 mo of age inside a inhabitants of CD34(-)KSL cells in addition to amongst unseparated bone marrow cells.
As well as, we detected one other compartment of repopulating cells, which differed from HSCs, amongst CD34(-)KSL cells of 18-mo-old mice. These repopulating cells confirmed much less differentiation potential towards lymphoid cells however retained self-renewal potential, as steered by secondary transplantation.
We suggest that HSCs progressively accumulate with age, accompanied by cells with much less lymphoid differentiation potential, because of repeated self-renewal of HSCs.

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MAGIC REDâ„¢ CASPASE-3/7 KIT (OKSA11289)

OKSA11289 100 Tests
EUR 1328
Description: Description of target: MAGIC REDâ„¢ CASPASE-3/7 KIT;Species reactivity: Human;Application: IF;Assay info: ;Sensitivity:

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K2053-100 100 assays
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1175-3
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CaspGLOW Red Active Caspase 3 Staining Kit

55R-1298 25 assays
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Description: Caspase 3 Staining Kit for use activity in the research laboratory

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CaspGLOW? Red Active Caspase-3 Staining Kit

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Red Active Caspase Staining Kit

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Caspase-3 DEVD-R110 Fluorometric HTS Assay Kit

30009-3 1KIT
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EUR 296
Description: Caspase Staining Kit for use activity in the research laboratory

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Red Active Caspase-8 Staining Kit

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Red Active Caspase-9 Staining Kit

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Red Active Caspase-9 Staining Kit

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MAGic Clamp

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MAGic Clamp

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MAGic Clamp

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EUR 201.59

MAGic Clamp

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EUR 154.61

MAGic Clamp

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EUR 146.78

MAGic Clamp

H1000-MR-TSR 1 PC
EUR 99.8

Caspase-8 Inhibitor, Q-IETD-OPh

1176-3
EUR 686

Caspase-9 Inhibitor, Q-LEHD-OPh

1177-3
EUR 675

Anti-Caspase-8 Monoclonal Antibody (5F7)

M00042-3 100ug
EUR 420
Description: Mouse Monoclonal Caspase-8 Antibody (5F7). Validated in IP, IHC, WB and tested in Human.

Anti-Caspase-9 Rabbit Monoclonal Antibody

M00080-3 100ug/vial
EUR 397
Description: Rabbit Monoclonal Caspase-9 Antibody. Validated in IP, IF, IHC, WB and tested in Human.

CaspGLOW Red Active Caspase 8 Staining Kit

55R-1299 25 assays
EUR 296
Description: Caspase 8 Staining Kit for use activity in the research laboratory

CaspGLOW Red Active Caspase 9 Staining Kit

55R-1300 25 assays
EUR 296
Description: Caspase 9 Staining Kit for use activity in the research laboratory

CaspGLOW? Red Active Caspase-8 Staining Kit

K198-100
EUR 523

CaspGLOW? Red Active Caspase-8 Staining Kit

K198-25
EUR 240

CaspGLOW? Red Active Caspase-9 Staining Kit

K199-100
EUR 517

CaspGLOW? Red Active Caspase-9 Staining Kit

K199-25
EUR 240

Amplite™ Fluorimetric Caspase 3/7 Assay Kit *Red Fluorescence*

13504 100 tests
EUR 350

Caspase-8 IETD-R110 Fluorometric HTS Assay Kit

30012-3 1KIT
EUR 1627
Description: Minimum order quantity: 1 unit of 1KIT

Caspase-3

MO15136 100 ug
EUR 466

Caspase-3

GT15044 100 ug
EUR 500

Cell Meter™ Caspase 3/7 Activity Apoptosis Assay Kit *Red Fluorescence*

22797 100 Tests
EUR 306

Cell Meter™ Live Cell Caspase 3/7 Binding Assay Kit *Red Fluorescence*

20101 25 Tests
EUR 306

Caspase 3 antibody

70R-49456 100 ul
EUR 244
Description: Purified Polyclonal Caspase 3 antibody

Caspase 3 antibody

70R-49457 100 ul
EUR 244
Description: Purified Polyclonal Caspase 3 antibody

Caspase 3 antibody

70R-49458 100 ul
EUR 244
Description: Purified Polyclonal Caspase 3 antibody

Caspase 3 antibody

70R-33463 100 ug
EUR 327
Description: Rabbit polyclonal Caspase 3 antibody

Caspase 3 Antibody

AF6311 200ul
EUR 304
Description: Caspase 3 Antibody detects endogenous levels of total Caspase 3.

Caspase 3 Antibody

ABF6311 100 ug
EUR 438

anti-Caspase 3

YF-PA10678 100 ul
EUR 403
Description: Rabbit polyclonal to Caspase 3

anti-Caspase 3

YF-PA10679 100 ug
EUR 403
Description: Rabbit polyclonal to Caspase 3

Caspase 3 protein

30R-2319 25 units
EUR 232
Description: Purified recombinant Human Caspase 3 protein

Caspase 3 protein

30R-2327 25 units
EUR 281
Description: Purified recombinant Human Caspase 3 protein

Caspase 3 protein

30R-2329 25 units
EUR 281
Description: Purified recombinant Human Caspase 3 protein

Caspase 3 protein

30R-2414 5 ug
EUR 486
Description: Purified recombinant Human Caspase 3 protein

Caspase 3 antibody

20R-1424 100 ug
EUR 781
Description: Rabbit polyclonal Caspase 3 antibody

Caspase 3 antibody

20R-1469 100 ug
EUR 651
Description: Rabbit polyclonal Caspase 3 antibody

Caspase 3 antibody

20R-2495 50 ug
EUR 281
Description: Rabbit polyclonal Caspase 3 antibody

Caspase 3 antibody

20R-CR013 50 ug
EUR 517
Description: Rabbit polyclonal Caspase 3 antibody

Caspase-3 Antibody

3138-100
EUR 359

Caspase-3 Antibody

3138-30T
EUR 146

Caspase 3 antibody

70R-11558 100 ug
EUR 645
Description: Rabbit polyclonal Caspase 3 antibody

Caspase 3 antibody

70R-11638 100 ug
EUR 470
Description: Rabbit polyclonal Caspase 3 antibody

Caspase 3 antibody

70R-31075 100 ug
EUR 327
Description: Rabbit polyclonal Caspase 3 antibody

Caspase 3 antibody

70R-31076 100 ug
EUR 327
Description: Rabbit polyclonal Caspase 3 antibody

Caspase 3 antibody

70R-31077 100 ug
EUR 327
Description: Rabbit polyclonal Caspase 3 antibody

Caspase 3 antibody

70R-13844 100 ug
EUR 322
Description: Affinity purified Rabbit polyclonal Caspase 3 antibody

Caspase 3 Antibody

DF6020 200ul
EUR 304
Description: CASP3 Antibody detects endogenous levels of total CASP3.

Caspase 3 Antibody

DF6879 200ul
EUR 304
Description: CASP3 Antibody detects endogenous levels of total CASP3.

Caspase 3 antibody

PAab10013 100 ug
EUR 386

Caspase 3 protein

E20-80020 20ug
EUR 408

Caspase 3 protein

E20-80021 20ug
EUR 408

Caspase 3 Antibody

21420-100ul 100ul
EUR 252

Caspase 3 Antibody

21420-50ul 50ul
EUR 187

Caspase-3 Antibody

25442-100ul 100ul
EUR 390

Caspase-3, active

RA15046 50 ug
EUR 383

Caspase-3 Antibody

48562-100ul 100ul
EUR 333

Caspase-3 Antibody

48562-50ul 50ul
EUR 239

Caspase-3 Antibody

48658-100ul 100ul
EUR 333

Caspase-3 Antibody

48658-50ul 50ul
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Cell Meter™ Caspase 8 Activity Apoptosis Assay Kit *Red Fluorescence*

22816 100 Tests
EUR 219

Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit *Red Fluorescence*

22817 100 Tests
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Caspase-3/CPP32/ Rat Caspase- 3/ CPP32 ELISA Kit

ELA-E0626r 96 Tests
EUR 886

 

 

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