Membrane Interactions of Cy3 and Cy5 Fluorophores and Their Effects on Membrane-Protein Dynamics
Natural fluorophores, equivalent to Cy3 and Cy5, have been extensively used as chemical labels to probe the construction and dynamics of membrane proteins. Though plenty of earlier research have reported on the opportunity of a few of the water-soluble fluorophores to work together with lipid bilayers, detailed fluorophore-lipid interactions and, extra importantly, the potential impact of such interactions on the pure dynamics of the labeled membrane proteins haven’t been properly studied.
We have now carried out a big set of all-atom molecular dynamics simulations using the extremely cell membrane mimetic mannequin to explain spontaneous partitioning of the fluorophores into lipid bilayers with totally different lipid compositions. Spontaneous membrane partitioning of Cy3 and Cy5 fluorophores captured in these simulations proceeds in two steps.
Electrostatic interplay between the fluorophores and the lipid headgroups facilitates the preliminary, quick membrane affiliation of the fluorophores, adopted by sluggish insertion of hydrophobic moieties into the lipid bilayer core. After the conversion of the ensuing membrane-bound programs to full-membrane representations, biased-exchange umbrella sampling simulations are carried out at no cost vitality calculations, revealing the next vitality barrier for partitioning into negatively charged (phosphatidylserine or phosphatidylcholine) membranes than purely zwitterionic (phosphatidylcholine or phosphatidylethanolamine) ones.
Moreover, the potential impact of fluorophore-lipid interactions on membrane proteins has been examined by covalently linking Cy5 to single- and multipass transmembrane helical proteins. Equilibrium simulations present sturdy position-dependent results of Cy5-tagging on the construction and pure dynamics of membrane proteins. Interactions between the tagged protein and Cy5 have been additionally noticed. Our outcomes counsel that fluorophore-lipid interactions can have an effect on the construction and dynamics of membrane proteins to varied extents, particularly in programs with larger structural flexibility.
Development and characterization of Cy3- or Cy5-conjugated hairpin pyrrole-imidazole polyamides binding to DNA within the nucleosome
Sequence-specific DNA-binding modules, N-methylpyrrole (Py)-N-methylimidazole-(Im) polyamides have been not too long ago conjugated with fluorophores, and a few of these conjugates could possibly be used for the detection of particular DNA sequences. On this examine, we synthesized two Py-Im polyamides 1 and a couple of, which work together with the 145-bp nucleosome positioning sequence 601.
We conjugated the cyanine dye Cy3 or Cy5 with 1 or 2. Within the absence of goal DNA, the fluorescent conjugate of a Py-Im polyamide had decrease fluorescence depth in contrast with Cy3 or Cy5 alone. Within the presence of both the goal DNA or the nucleosome, the fluorescence depth of the conjugates elevated. Moreover, we noticed a Förster resonance vitality switch between the Cy3-Py-Im polyamide and the Cy5-Py-Im polyamide on the nucleosome. These outcomes open up the chances that fluorescent conjugates of Py-Im polyamides can be utilized for characterization of the dynamic interactions inside protein-DNA complexes.
On-resin technique to label α-conotoxins: Cy5-RgIA, a potent α9α10 nicotinic acetylcholine receptor imaging probe.
In-solution conjugation is probably the most generally used technique to label peptides and proteins with fluorophores. Nonetheless, lack of site-specific management and excessive prices of fluorophores are recognised limitations of this method. Right here, we established facile entry to grams of Cy5-COOH through a two-step artificial route, demonstrated that Cy5 is steady to HF remedy and due to this fact appropriate with Boc-SPPS, and matched Cy5 to the N-terminus of α-conotoxin RgIA whereas nonetheless connected to the resin.
Folding of the two-disulfide containing Cy5-RgIA benefitted from the hydrophobic nature of Cy5 leading to solely the globular disulfide bond isomer. In distinction, wild-type α-RgIA folded into the inactive ribbon and bioactive globular isomer underneath the identical situations.
Labelled α-RgIA retained its potential to inhibit acetylcholine(100 μM)-evoked present reversibly with an IC50 of 5.zero nM (Hill coefficient = 1.7) for α-RgIA and an IC50 of 1.6 (Hill coefficient = 1.2) for Cy5-RgIA on the α9α10 nicotinic acetylcholine receptors (nAChRs) heterologeously expressed in Xenopus oocytes. Cy5-RgIA was then used to efficiently visualise nAChRs in RAW264.7 mouse macrophage cell line.
This work launched not solely a brand new and useful nAChR probe, but in addition a brand new versatile artificial technique that facilitates manufacturing of milligram to gram portions of fluorophore-labelled peptides at low value, which is commonly required for in vivo experiments. The technique is appropriate with Boc- and Fmoc-chemistry, permits for site-specific labelling of free amines anyplace within the peptide sequence, and can be used for the introduction of Cy3/Cy5 FRET pairs.
Ultrafast Excitation Switch in Cy5 DNA Photonic Wires Shows Dye Conjugation and Excitation Vitality Dependency.
DNA scaffolds allow base-pair-specific positioning of fluorescent molecules, permitting for nanometer-scale precision in controlling multidye interactions. Increasing on this idea, DNA-based molecular photonic wires (MPWs) permit for mild harvesting and directional propagation of photonic vitality on the nanometer scale. The most typical MPW examples exploit Förster resonance vitality switch (FRET), and FRET between the identical dye species (HomoFRET) was not too long ago proven to extend the gap and effectivity at which MPWs can perform.
Though elevated proximity between adjoining fluorophores can be utilized to extend the vitality switch effectivity, FRET assumptions break down as the gap between the dye molecules turns into similar to their measurement (∼2 nm). Right here we evaluate dye conjugation with single versus dimer Cy5 dye repeats as HomoFRET MPW elements on a double-crossover DNA scaffold.
At room temperature (RT) underneath low-light situations, end-labeled uncoupled dye molecules present optimum switch, whereas the Cy5 dimers present ultrafast (<100 ps) nonradiative decay that severely limits their performance. Of explicit curiosity is the statement that via elevated excitation fluence in addition to cryogenic temperatures, the dimeric MPW reveals suppression of the ultrafast decay, demonstrating fluorescence lifetimes much like the one Cy5 MPWs.
C21orf62 Antibody |
MBS9631185-02mL |
MyBiosource |
0.2mL |
EUR 305 |
C21orf62 Antibody |
MBS9631185-5x02mL |
MyBiosource |
5x0.2mL |
EUR 1220 |
C21orf62 Antibody |
MBS9633568-1mg |
MyBiosource |
1mg |
EUR 375 |
C21orf62 Antibody |
MBS9633568-5x1mg |
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C21orf62-AS1 Antibody |
20-abx306803 |
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C21orf62-AS1 Antibody |
1-CSB-PA613779LA01HU |
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Description: A polyclonal antibody against C21orf62-AS1. Recognizes C21orf62-AS1 from Human. This antibody is Unconjugated. Tested in the following application: ELISA |
C21orf62-AS1 Antibody |
abx306803-100g |
Abbexa |
100 µg |
EUR 362.5 |
C21orf62-AS1 Antibody |
abx306803-20g |
Abbexa |
20 µg |
EUR 162.5 |
C21orf62-AS1 Antibody |
abx306803-50g |
Abbexa |
50 µg |
EUR 250 |
C21orf62-AS1 Antibody |
MBS7001526-005mg |
MyBiosource |
0.05mg |
EUR 190 |
C21orf62-AS1 Antibody |
MBS7001526-01mg |
MyBiosource |
0.1mg |
EUR 270 |
C21orf62-AS1 Antibody |
MBS7001526-5x01mg |
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EUR 1205 |
C21ORF62 Polyclonal Antibody |
MBS9239022-01mL |
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0.1mL |
EUR 415 |
C21ORF62 Polyclonal Antibody |
MBS9239022-5x01mL |
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C21orf62-AS1 Antibody (HRP) |
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C21orf62-AS1 Antibody (HRP) |
abx306804-100g |
Abbexa |
100 µg |
EUR 362.5 |
C21orf62-AS1 Antibody (HRP) |
abx306804-20g |
Abbexa |
20 µg |
EUR 162.5 |
C21orf62-AS1 Antibody (HRP) |
abx306804-50g |
Abbexa |
50 µg |
EUR 250 |
C21orf62 antibody - middle region |
MBS3207949-01mL |
MyBiosource |
0.1mL |
EUR 455 |
C21orf62 antibody - middle region |
MBS3207949-5x01mL |
MyBiosource |
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EUR 1995 |
C21orf62-AS1 Antibody (FITC) |
20-abx306805 |
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C21orf62-AS1 Antibody (FITC) |
abx306805-100g |
Abbexa |
100 µg |
EUR 362.5 |
C21orf62-AS1 Antibody (FITC) |
abx306805-20g |
Abbexa |
20 µg |
EUR 162.5 |
C21orf62-AS1 Antibody (FITC) |
abx306805-50g |
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50 µg |
EUR 250 |
C21orf62-AS1 Antibody (Biotin) |
20-abx306806 |
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C21orf62-AS1 Antibody (Biotin) |
abx306806-100g |
Abbexa |
100 µg |
EUR 362.5 |
C21orf62-AS1 Antibody (Biotin) |
abx306806-20g |
Abbexa |
20 µg |
EUR 162.5 |
C21orf62-AS1 Antibody (Biotin) |
abx306806-50g |
Abbexa |
50 µg |
EUR 250 |
C21orf62-AS1 Polyclonal Antibody |
A64662 |
EpiGentek |
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Polyclonal C21orf62 antibody - middle region |
APR00970G |
Leading Biology |
0.05mg |
EUR 633.6 |
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human C21orf62 - middle region. This antibody is tested and proven to work in the following applications: |
C21orf62-AS1 Antibody, HRP conjugated |
1-CSB-PA613779LB01HU |
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Description: A polyclonal antibody against C21orf62-AS1. Recognizes C21orf62-AS1 from Human. This antibody is HRP conjugated. Tested in the following application: ELISA |
C21orf62-AS1 Antibody; HRP conjugated |
MBS7000751-005mg |
MyBiosource |
0.05mg |
EUR 190 |
C21orf62-AS1 Antibody; HRP conjugated |
MBS7000751-01mg |
MyBiosource |
0.1mg |
EUR 270 |
C21orf62-AS1 Antibody; HRP conjugated |
MBS7000751-5x01mg |
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5x0.1mg |
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C21orf62-AS1 Antibody, FITC conjugated |
1-CSB-PA613779LC01HU |
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Description: A polyclonal antibody against C21orf62-AS1. Recognizes C21orf62-AS1 from Human. This antibody is FITC conjugated. Tested in the following application: ELISA |
C21orf62-AS1 Antibody; FITC conjugated |
MBS7000588-005mg |
MyBiosource |
0.05mg |
EUR 190 |
C21orf62-AS1 Antibody; FITC conjugated |
MBS7000588-01mg |
MyBiosource |
0.1mg |
EUR 270 |
C21orf62-AS1 Antibody; FITC conjugated |
MBS7000588-5x01mg |
MyBiosource |
5x0.1mg |
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C21orf62-AS1 Antibody, Biotin conjugated |
1-CSB-PA613779LD01HU |
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Description: A polyclonal antibody against C21orf62-AS1. Recognizes C21orf62-AS1 from Human. This antibody is Biotin conjugated. Tested in the following application: ELISA |
C21orf62-AS1 Antibody; Biotin conjugated |
MBS7001693-005mg |
MyBiosource |
0.05mg |
EUR 190 |
C21orf62-AS1 Antibody; Biotin conjugated |
MBS7001693-01mg |
MyBiosource |
0.1mg |
EUR 270 |
C21orf62-AS1 Antibody; Biotin conjugated |
MBS7001693-5x01mg |
MyBiosource |
5x0.1mg |
EUR 1205 |
ARP45868_P050-25UL - C21orf62 Antibody - middle region |
ARP45868_P050-25UL |
Aviva Systems Biology |
25ul |
EUR 99 |
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ARP45868_P050 - C21orf62 antibody - middle region (ARP45868_P050) |
ARP45868_P050 |
Aviva Systems Biology |
100ul |
EUR 389 |
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C21orf62-AS1 Polyclonal Antibody, HRP Conjugated |
A64663 |
EpiGentek |
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C21orf62-AS1 Polyclonal Antibody, FITC Conjugated |
A64664 |
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C21orf62-AS1 Polyclonal Antibody, Biotin Conjugated |
A64665 |
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Recombinant Human Uncharacterized protein C21orf62 (C21orf62) |
MBS1428897-002mgBaculovirus |
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0.02mg(Baculovirus) |
EUR 1135 |
Recombinant Human Uncharacterized protein C21orf62 (C21orf62) |
MBS1428897-002mgEColi |
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0.02mg(E-Coli) |
EUR 725 |
Recombinant Human Uncharacterized protein C21orf62 (C21orf62) |
MBS1428897-002mgYeast |
MyBiosource |
0.02mg(Yeast) |
EUR 890 |
Recombinant Human Uncharacterized protein C21orf62 (C21orf62) |
MBS1428897-01mgEColi |
MyBiosource |
0.1mg(E-Coli) |
EUR 870 |
Recombinant Human Uncharacterized protein C21orf62 (C21orf62) |
MBS1428897-01mgYeast |
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0.1mg(Yeast) |
EUR 1045 |
C21orf62 siRNA |
20-abx909758 |
Abbexa |
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Human Uncharacterized protein C21orf62, C21orf62 ELISA KIT |
ELI-33580h |
Nova Lifetech |
96tests |
EUR 696 |
Human Uncharacterized protein C21orf62 (C21orf62) ELISA Kit |
abx509209-96tests |
Abbexa |
96 tests |
EUR 687.5 |
Human Uncharacterized protein C21orf62, C21orf62 ELISA Kit |
MBS9325137-10x96StripWells |
MyBiosource |
10x96-Strip-Wells |
EUR 6725 |
Human Uncharacterized protein C21orf62, C21orf62 ELISA Kit |
MBS9325137-48StripWells |
MyBiosource |
48-Strip-Wells |
EUR 550 |
Human Uncharacterized protein C21orf62, C21orf62 ELISA Kit |
MBS9325137-5x96StripWells |
MyBiosource |
5x96-Strip-Wells |
EUR 3420 |
Human Uncharacterized protein C21orf62, C21orf62 ELISA Kit |
MBS9325137-96StripWells |
MyBiosource |
96-Strip-Wells |
EUR 765 |
C21orf62 siRNA (Human) |
MBS8214308-15nmol |
MyBiosource |
15nmol |
EUR 405 |
C21orf62 siRNA (Human) |
MBS8214308-30nmol |
MyBiosource |
30nmol |
EUR 565 |
C21orf62 siRNA (Human) |
MBS8214308-5x30nmol |
MyBiosource |
5x30nmol |
EUR 2450 |
C21ORF62 Blocking Peptide |
33R-8879 |
Fitzgerald |
100 ug |
EUR 119 |
|
Description: A synthetic peptide for use as a blocking control in assays to test for specificity of C21orf62 antibody, catalog no. 70R-5266 |
Human C21orf62 shRNA Plasmid |
20-abx961112 |
Abbexa |
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Human C21ORF62 Protein Lysate |
MBS138368-002mg |
MyBiosource |
0.02mg |
EUR 365 |
Human C21ORF62 Protein Lysate |
MBS138368-5x002mg |
MyBiosource |
5x0.02mg |
EUR 1410 |
C21orf62 Peptide - middle region |
MBS3232915-01mg |
MyBiosource |
0.1mg |
EUR 180 |
C21orf62 Peptide - middle region |
MBS3232915-5x01mg |
MyBiosource |
5x0.1mg |
EUR 730 |
C21orf62 Recombinant Protein (Human) |
RP049810 |
ABM |
100 ug |
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Carrier-free (BSA/glycerol-free) C21orf62 mouse monoclonal antibody,clone OTI9C8 |
CF808845 |
Origene Technologies GmbH |
100 µg |
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Carrier-free (BSA/glycerol-free) C21orf62 mouse monoclonal antibody,clone OTI4A6 |
CF808847 |
Origene Technologies GmbH |
100 µg |
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Carrier-free (BSA/glycerol-free) C21orf62 mouse monoclonal antibody,clone OTI5B5 |
CF808848 |
Origene Technologies GmbH |
100 µg |
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C21orf62 (NM_001162495) Human Recombinant Protein |
PROTQ9NYP8 |
BosterBio |
20ug |
EUR 1249 |
Description: Purified recombinant protein of Homo sapiens chromosome 21 open reading frame 62 (C21orf62), transcript variant 1. |
C21orf62 (untagged)-Human chromosome 21 open reading frame 62 (C21orf62) transcript variant 1 |
SC327401 |
Origene Technologies GmbH |
10 µg |
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C21orf62 (untagged)-Human chromosome 21 open reading frame 62 (C21orf62) transcript variant 3 |
SC327402 |
Origene Technologies GmbH |
10 µg |
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C21orf62 (untagged)-Human chromosome 21 open reading frame 62 (C21orf62), transcript variant 2 |
SC317354 |
Origene Technologies GmbH |
10 µg |
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C21orf62 ORF Vector (Human) (pORF) |
ORF016604 |
ABM |
1.0 ug DNA |
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Human C21orf62 knockout cell line |
ABC-KH1974 |
AcceGen |
1 vial |
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Description: Human C21orf62 knockout cell line is HEK293/HeLa cell line, edited by CRISPR/Cas9 technology. |
Human C21orf62 knockdown cell line |
ABC-KD1974 |
AcceGen |
1 vial |
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Description: Human C21orf62 knockdown cell line is engineered by our optimized transduction of the specific shRNA with lentivirus. Knockdown levels are determined via qRT-PCR. Gentaur offers generation of stable knockdown (RNAi) cell lines expressing shRNAs targeting genes of your interest. |
Human C21ORF62 Protein Lysate 20ug |
IHUC21ORF62PLLY20UG |
Innovative research |
each |
EUR 213 |
|
Description: Human C21ORF62 Protein Lysate 20ug |
C21orf62 (GFP-tagged) - Human chromosome 21 open reading frame 62 (C21orf62), transcript variant 2 |
RG222481 |
Origene Technologies GmbH |
10 µg |
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C21orf62 (GFP-tagged) - Human chromosome 21 open reading frame 62 (C21orf62), transcript variant 1 |
RG228766 |
Origene Technologies GmbH |
10 µg |
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C21orf62 (GFP-tagged) - Human chromosome 21 open reading frame 62 (C21orf62), transcript variant 3 |
RG228767 |
Origene Technologies GmbH |
10 µg |
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C21orf62 Over-expression Lysate Product |
GWB-299154 |
GenWay Biotech |
0.1 mg |
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Carrier-free (BSA/glycerol-free) C21orf62 mouse monoclonal antibody, clone OTI6C2 (formerly 6C2) |
CF808826 |
Origene Technologies GmbH |
100 µg |
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Carrier-free (BSA/glycerol-free) C21orf62 mouse monoclonal antibody, clone OTI1B8 (formerly 1B8) |
CF808837 |
Origene Technologies GmbH |
100 µg |
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C21orf62 (NM_001162495) Human Over-expression Lysate |
LS056245 |
BosterBio |
100ug |
EUR 628 |
|
Description: Transient overexpression lysate of chromosome 21 open reading frame 62 (C21orf62) |
C21orf62 (Myc-DDK-tagged)-Human chromosome 21 open reading frame 62 (C21orf62), transcript variant 2 |
RC222481 |
Origene Technologies GmbH |
10 µg |
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C21orf62 (Myc-DDK-tagged)-Human chromosome 21 open reading frame 62 (C21orf62), transcript variant 1 |
RC228766 |
Origene Technologies GmbH |
10 µg |
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C21orf62 (Myc-DDK-tagged)-Human chromosome 21 open reading frame 62 (C21orf62), transcript variant 3 |
RC228767 |
Origene Technologies GmbH |
10 µg |
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C21orf62 sgRNA CRISPR Lentivector set (Human) |
K0340701 |
ABM |
3 x 1.0 ug |
EUR 406.8 |
C21orf62 3'UTR GFP Stable Cell Line |
TU053265 |
ABM |
1.0 ml |
EUR 1672.8 |
C21orf62 Protein Vector (Human) (pPM-C-HA) |
PV066415 |
ABM |
500 ng |
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C21orf62 3'UTR Luciferase Stable Cell Line |
TU003265 |
ABM |
1.0 ml |
EUR 1672.8 |
C21orf62 Protein Vector (Human) (pPB-C-His) |
PV066413 |
ABM |
500 ng |
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C21orf62 Protein Vector (Human) (pPB-N-His) |
PV066414 |
ABM |
500 ng |
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C21orf62 Protein Vector (Human) (pPM-C-His) |
PV066416 |
ABM |
500 ng |
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Recombinant Mouse Uncharacterized protein C21orf62 homolog |
MBS1334010-002mgBaculovirus |
MyBiosource |
0.02mg(Baculovirus) |
EUR 1120 |
Recombinant Mouse Uncharacterized protein C21orf62 homolog |
MBS1334010-002mgEColi |
MyBiosource |
0.02mg(E-Coli) |
EUR 740 |
Recombinant Mouse Uncharacterized protein C21orf62 homolog |
MBS1334010-002mgYeast |
MyBiosource |
0.02mg(Yeast) |
EUR 900 |
Recombinant Mouse Uncharacterized protein C21orf62 homolog |
MBS1334010-01mgEColi |
MyBiosource |
0.1mg(E-Coli) |
EUR 885 |
Recombinant Mouse Uncharacterized protein C21orf62 homolog |
MBS1334010-01mgYeast |
MyBiosource |
0.1mg(Yeast) |
EUR 1055 |
C21orf62 Protein Vector (Human) (pPB-His-MBP) |
PV330542 |
ABM |
500 ng |
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C21orf62 Protein Vector (Human) (pPB-His-GST) |
PV330543 |
ABM |
500 ng |
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C21orf62 sgRNA CRISPR Lentivector (Human) (Target 1) |
K0340702 |
ABM |
1.0 ug DNA |
EUR 184.8 |
C21orf62 sgRNA CRISPR Lentivector (Human) (Target 2) |
K0340703 |
ABM |
1.0 ug DNA |
EUR 184.8 |
C21orf62 sgRNA CRISPR Lentivector (Human) (Target 3) |
K0340704 |
ABM |
1.0 ug DNA |
EUR 184.8 |
Human Putative uncharacterized protein C21orf49 (C21orf62-AS1) ELISA Kit |
abx509204-96tests |
Abbexa |
96 tests |
EUR 687.5 |
C21orf62 Protein Vector (Human) (pPM-N-D-C-HA) |
PV330544 |
ABM |
500 ng |
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C21orf62 Protein Vector (Human) (pPM-N-D-C-His) |
PV330545 |
ABM |
500 ng |
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Lenti ORF particles, C21orf62 (mGFP-tagged) - Human chromosome 21 open reading frame 62 (C21orf62), transcript variant 2, 200ul, >10^7 TU/mL |
RC222481L2V |
Origene Technologies GmbH |
200 µl |
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Lenti ORF particles, C21orf62 (mGFP-tagged) - Human chromosome 21 open reading frame 62 (C21orf62), transcript variant 2, 200ul, >10^7 TU/mL |
RC222481L4V |
Origene Technologies GmbH |
200 µl |
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Lenti ORF particles, C21orf62 (mGFP-tagged) - Human chromosome 21 open reading frame 62 (C21orf62), transcript variant 1, 200ul, >10^7 TU/mL |
RC228766L2V |
Origene Technologies GmbH |
200 µl |
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This work factors to the complicated dynamic capabilities of dye-based nanophotonic networks, the place dye positioning and interactions can develop into essential, and could possibly be used to increase the lengths and complexities of such dye-DNA gadgets, enabling multiparameter nanophotonic circuitry.
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