Isolation and Characterization of Extracellular Vesicles from Cell Culture Conditioned Medium for Immunological Studies

Extracellular vesicles (EVs) are small, membranous particles which have just lately emerged as one a very powerful mediators of intercellular communication. They will comprise quite a lot of proteins, lipids, and nucleic acids and thus are answerable for modulation of a number of organic processes, together with immune response and regulation of immune cells. Immunomodulatory exercise of various EVs might be reliably assessed utilizing EVs remoted from cell tradition conditioned medium and added to in vitro or ex vivo cultures of immune cells. This text describes protocols for isolation of EVs from cell tradition supernatants by differential ultracentrifugation and density gradient centrifugation. It additionally offers instruments and protocols that allow characterization and validation of remoted particles, in addition to evaluation of interactions between EVs of curiosity and totally different subpopulations of human immune cells.

2020 Wiley Periodicals LLC. Fundamental Protocol 1: Isolation of extracellular vesicles by differential ultracentrifugation Fundamental Protocol 2: Isolation of extracellular vesicles by density gradient centrifugation Help Protocol 1: Imaging of extracellular vesicles utilizing transmission electron microscopy Help Protocol 2: Detection of extracellular vesicle protein markers by Western blotting Help Protocol 3: Measurement and counting of extracellular vesicles by nanoparticle monitoring evaluation Fundamental Protocol 3: Evaluation of extracellular vesicle uptake or affiliation by totally different subpopulations of lymphocytes in vitro.

 

Simultaneous Quantification of Chosen Kynurenines Analyzed by Liquid Chromatography-Mass Spectrometry in Medium Collected From Most cancers Cell Cultures

The kynurenine pathway and the tryptophan catabolites known as kynurenines have obtained elevated consideration for his or her involvement in immune regulation and most cancers biology. An in vitro cell tradition assay is commonly used to be taught in regards to the contribution of various tryptophan catabolites in a illness mechanism and for testing therapeutic methods. Cell tradition medium that’s wealthy in secreted metabolites and signaling molecules displays the standing of tryptophan metabolism and different mobile occasions. New protocols for the dependable quantification of a number of kynurenines within the complicated cell tradition medium are desired to permit for a dependable and fast evaluation of a number of samples.
This may be completed with liquid chromatography coupled with mass spectrometry. This highly effective method is employed in lots of scientific and analysis laboratories for the quantification of metabolites and can be utilized for measuring kynurenines. Offered right here is the usage of liquid chromatography coupled with single quadrupole mass spectrometry (LC-SQ) for the simultaneous willpower of 4 kynurenines, i.e., kynurenine, 3-hydroxykynurenine, 3-hydroxyanthranilic, and xanthurenic acid within the medium collected from in vitro cultured most cancers cells. SQ detector is easy to make use of and cheaper in comparison with different mass spectrometers. Within the SQ-MS evaluation, a number of ions from the pattern are generated and separated based on their particular mass-to-charge ratio (m/z), adopted by the detection utilizing a Single Ion Monitoring (SIM) mode.
This paper attracts the eye on some great benefits of the reported technique and signifies some weak factors. It’s centered on vital parts of LC-SQ evaluation together with pattern preparation together with chromatography and mass spectrometry evaluation. The standard management, technique calibration situations and matrix impact points are additionally mentioned. We described a easy software of 3-nitrotyrosine as one analog customary for all goal analytes. As confirmed by experiments with human ovary and breast most cancers cells, the proposed LC-SQ technique generates dependable outcomes and might be additional utilized to different in vitro mobile fashions.

Direct counting of exosomes in a cell tradition medium utilizing neither isolation nor preconcentration

Exosomes are anticipated to be biomarkers of most cancers since they comprise details about the cells that excrete them. On this examine we developed a way to depend the exosomes secreted from most cancers cells in a tradition medium with out the necessity for isolation and/or preconcentration. This detection system consists of a sq. capillary on which a laser beam is concentrated in a sheet form by way of the usage of two cylindrical lenses. A fluorescently labeled anti-CD63 antibody is used to mark the exosomes which can be then flowed into the sq. capillary.

Chikungunya virus RT PCR kit

RTq-H525-150R 150T
EUR 2144.4

Chikungunya virus RT PCR kit

RTq-H525-50R 50T
EUR 1155.6

Truenat™ Chip-based Real Time PCR Test Chikungunya

Chikungunya each
EUR 18

CHIKUNGUNYA DETECTION KIT (REAL TIME PRO

MBPCR112-10R 1 unit
EUR 115.82
Description: CHIKUNGUNYA DETECTION KIT (REAL TIME PRO

CHIKUNGUNYA DETECTION KIT (REAL TIME PRO

MBPCR112-50R 1 unit
EUR 505.92
Description: CHIKUNGUNYA DETECTION KIT (REAL TIME PRO

Chikungunya virus PCR kit

PCR-H525-48R 50T
EUR 987.6

Chikungunya virus PCR kit

PCR-H525-96R 100T
EUR 1335.6

Truenat™ Chip-based Real Time PCR Test Dengue/ Chikungunya

Dengue/_Chikungunya each
EUR 20

genesig Std Real-time PCR detection kit for Chikungunya Vir

Z-Path-CHIKV-std 150 tests
EUR 602
Description: CHIKV

Chikungunya virus One-Step PCR kit

Oneq-H525-100R 100T
EUR 1932

Chikungunya virus One-Step PCR kit

Oneq-H525-150R 150T
EUR 2646

Chikungunya virus One-Step PCR kit

Oneq-H525-50R 50T
EUR 1410

Chikungunya virus

PCR-H525-PCRH52548R PCR-H525-48R
EUR 485

Chikungunya virus

PCR-H525-PCRH52596R PCR-H525-96R
EUR 685

Chikungunya virus

Oneq-H525-OneqH525100R Oneq-H525-100R
EUR 1028

Chikungunya virus

Oneq-H525-OneqH525150R Oneq-H525-150R
EUR 1438

Chikungunya virus

Oneq-H525-OneqH52550R Oneq-H525-50R
EUR 728

ViPrimePLUS Chikungunya Virus RT-qPCR Kit, 150rxns

QM5004 each Ask for price

Chikungunya virus lysate

NAT41557-100 0.1
EUR 512.85
Description: Purified Chikungunya virus lysate.

Chikungunya Virus IgG ELISA Kit

ESR148G 96 Tests
EUR 372

Chikungunya Virus IgM ELISA Kit

ESR148M 96 Tests
EUR 372

Chikungunya Virus Antibody

abx021574-1mg 1 mg
EUR 1296

Chikungunya Virus Antibody

abx021575-1mg 1 mg
EUR 1296

Chikungunya Virus Antibody

abx021576-1mg 1 mg
EUR 1296

Chikungunya virus antibody

10-2716 500 ug
EUR 295
Description: Mouse monoclonal Chikungunya virus envelope antibody

Chikungunya virus antibody

10-2717 500 ug
EUR 295
Description: Mouse monoclonal Chikungunya virus envelope antibody

Chikungunya virus antibody

10-2718 500 ug
EUR 300
Description: Mouse monoclonal Chikungunya virus envelope antibody

Chikungunya virus antibody

10-2719 250 ug
EUR 492
Description: Mouse monoclonal Chikungunya virus antibody

Chikungunya virus antibody

10-2720 250 ug
EUR 590
Description: Mouse monoclonal Chikungunya virus antibody

Chikungunya Virus Antibody

abx021574-400l 400 µl Ask for price

Chikungunya Virus Antibody

abx021574-80l 80 µl
EUR 1250

Chikungunya Virus Antibody

abx021575-400l 400 µl Ask for price

Chikungunya Virus Antibody

abx021575-80l 80 µl
EUR 1250

Chikungunya Virus Antibody

abx021576-400l 400 µl Ask for price

Chikungunya Virus Antibody

abx021576-80l 80 µl
EUR 1250

QPCR Kit RNA Chikungunya virus - EACH

MOL6850 EACH
EUR 1176.51

Chikungunya virus E1 protein

30-1942 100 ug
EUR 1500
Description: Recombinant Chikungunya virus E1 protein - wild type

Chikungunya Virus IgG capture

GWB-C324EF 1x96 Assays Ask for price

Human Chikungunya Virus (CHIKV) ELISA Kit

SL3467Hu 96 Tests
EUR 468

Chikungunya virus gpE2 protein

30-1939 100 ug
EUR 1560
Description: Recombinant Chikungunya virus Glycoprotein E2 protein - wild type

Chikungunya virus gpE1 protein

30-1940 100 ug
EUR 1560
Description: Recombinant Chikungunya virus Glycoprotein E1 protein - wild type

Chikungunya virus gpE1 protein

30-1944 100 ug
EUR 1184
Description: Recombinant Chikungunya virus Glycoprotein E1 protein - wild type

genesig Easy kit for Chikungunya Virus

Z-Path-CHIKV-EASY 50 tests
EUR 410
Description: CHIKV

Eukaryotic Chikungunya Virus (CHIKV)

EPX256Ge41 10ug
EUR 240

Eukaryotic Chikungunya Virus (CHIKV)

RPU59037-100ug 100ug
EUR 950.4

Eukaryotic Chikungunya Virus (CHIKV)

RPU59037-1mg 1mg
EUR 3744

Eukaryotic Chikungunya Virus (CHIKV)

RPU59037-50ug 50ug
EUR 594

Recombinant Chikungunya Virus E1

P1113-10 each
EUR 614.4

Recombinant Chikungunya Virus (CHIKV)

RPU56069-100ug 100ug
EUR 871.2

Recombinant Chikungunya Virus (CHIKV)

RPU56069-1mg 1mg
EUR 3432

Recombinant Chikungunya Virus (CHIKV)

RPU56069-50ug 50ug
EUR 528

Recombinant Chikungunya Virus (CHIKV)

RPX256Ge01 10ug
EUR 220

Recombinant Chikungunya Virus (CHIKV)

4-RPX256Ge01
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  • 1 mg
  • 200 ug
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  • 50ug
  • 5 mg
Description: Recombinant Pan-species Chikungunya Virus expressed in: E.coli

Recombinant Chikungunya Virus VLP

DAGA-3014 100ug
EUR 1622.4

Chikungunya Virus Envelope Antibody

abx024072-1mg 1 mg
EUR 1161.6

Chikungunya Virus Envelope Antibody

abx024073-1mg 1 mg
EUR 1161.6

Chikungunya Virus Envelope Antibody

abx024074-1mg 1 mg
EUR 1161.6

Chikungunya Virus Envelope Antibody

abx024075-1mg 1 mg
EUR 1161.6

Chikungunya Virus Envelope Antibody

abx024076-1mg 1 mg
EUR 1161.6

Chikungunya Virus Envelope Antibody

abx024074-400l 400 µl Ask for price

Chikungunya Virus Envelope Antibody

abx024074-80l 80 µl
EUR 1037.5

Chikungunya Virus Envelope Antibody

abx024075-400l 400 µl Ask for price

Chikungunya Virus Envelope Antibody

abx024075-80l 80 µl
EUR 1037.5

Chikungunya Virus Envelope Antibody

abx024076-400l 400 µl Ask for price

Chikungunya Virus Envelope Antibody

abx024076-80l 80 µl
EUR 1037.5

OVA Conjugated Chikungunya Virus (CHIKV)

CPX256Ge21 10ug
EUR 220

OVA Conjugated Chikungunya Virus (CHIKV)

4-CPX256Ge21
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  • 100 ug
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  • 5 mg
Description: Recombinant Pan-species Chikungunya Virus expressed in: Protein conjugation

OVA Conjugated Chikungunya Virus (CHIKV)

RPU56068-100ug 100ug
EUR 871.2

OVA Conjugated Chikungunya Virus (CHIKV)

RPU56068-1mg 1mg
EUR 3432

OVA Conjugated Chikungunya Virus (CHIKV)

RPU56068-50ug 50ug
EUR 605

Mouse antibody for Chikungunya virus

3584 100 ug
EUR 386.26
Description: This is purified Mouse monoclonal antibody against Chikungunya virus for WB, ELISA.

Mouse Anti Chikungunya Virus VLP (D7)

MAB12385-100 0.1
EUR 341.9
Description: Mouse monoclonal antibody specific for Chikungunya virus VLP (clone D7)

Mouse Anti Chikungunya Virus VLP (D7)

MAB12385-200 0.2
EUR 494.42
Description: Mouse monoclonal antibody specific for Chikungunya virus VLP (clone D7)

Mouse Anti Chikungunya Virus VLP (D7)

MAB12385-500 0.5
EUR 1454.36
Description: Mouse monoclonal antibody specific for Chikungunya virus VLP (clone D7)

Mouse Anti Chikungunya Virus VLP (AG7)

MAB12313-100 0.1
EUR 276.84
Description: Mouse monoclonal antibody specific for Chikungunya virus VLP (clone AG7)

Mouse Anti Chikungunya Virus VLP (AG7)

MAB12313-500 0.5
EUR 692.74
Description: Mouse monoclonal antibody specific for Chikungunya virus VLP (clone AG7)

Mouse Anti Chikungunya Virus VLP (EB6)

MAB12314-100 0.1
EUR 276.84
Description: Mouse monoclonal antibody specific for Chikungunya virus VLP (clone EB6)

Mouse Anti Chikungunya Virus VLP (EB6)

MAB12314-500 0.5
EUR 692.74
Description: Mouse monoclonal antibody specific for Chikungunya virus VLP (clone EB6)

OONB00094-100UG - Chikungunya virus VLP

OONB00094-100UG 100ug
EUR 979

OONB00094-500UG - Chikungunya virus VLP

OONB00094-500UG 500ug
EUR 3659

Human Chikungunya Virus IgG (CHIKV-IgG) ELISA Kit

abx054994-96tests 96 tests
EUR 801.6

Human Chikungunya Virus IgM (CHIKV-IgM) ELISA Kit

abx055296-96tests 96 tests
EUR 801.6

Human chikungunya virus IgM(CHIKV-IgM) Elisa Kit

EK711693 96 Wells
EUR 0.19

Human chikungunya virus IgG (CHIKV-IgG) Elisa Kit

EK700016 96 Wells
EUR 0.15

Human chikungunya virus IgG (CHIKV-IgG) ELISA Kit

SL2049Hu 96 Tests
EUR 468

Human chikungunya virus IgM (CHIKV-IgM) ELISA Kit

SL2160Hu 96 Tests
EUR 468

Human Chikungunya Virus IgG (CHIKV-IgG) ELISA Kit

abx054994-100g 100 µg Ask for price

Human Chikungunya Virus IgG (CHIKV-IgG) ELISA Kit

abx054994-10g 10 µg
EUR 493.75

Human Chikungunya Virus IgG (CHIKV-IgG) ELISA Kit

abx054994-50g 50 µg Ask for price

Human Chikungunya Virus IgM (CHIKV-IgM) ELISA Kit

abx055296-100g 100 µg Ask for price

Human Chikungunya Virus IgM (CHIKV-IgM) ELISA Kit

abx055296-10g 10 µg
EUR 493.75

Human Chikungunya Virus IgM (CHIKV-IgM) ELISA Kit

abx055296-50g 50 µg Ask for price

Chikungunya Virus (CHIKV) Monoclonal Antibody

CAU30851-100ul 100ul
EUR 436.8

Chikungunya Virus (CHIKV) Monoclonal Antibody

CAU30851-200ul 200ul
EUR 546

Chikungunya Virus IgM μ-capture ELISA

GWB-7CF864 1x96 Assays Ask for price

Chikungunya Virus VLP (E1 E2 C protein)

CHIKV-VLP-100 0.1
EUR 710.6
Description: Recombinant Chikungunya Virus-Like Particles produced in 293 human cells. Chikungunya structural proteins are transiently expressed to form a particle comprising E1, E2 and capsid protein, without the non-structural proteins or genome. Resultant particles are totally non-infectious.

Chikungunya Virus VLP (E1 E2 C protein)

CHIKV-VLP-1000 1.0
EUR 4748.34
Description: Recombinant Chikungunya Virus-Like Particles produced in 293 human cells. Chikungunya structural proteins are transiently expressed to form a particle comprising E1, E2 and capsid protein, without the non-structural proteins or genome. Resultant particles are totally non-infectious. This product is prepared from HEK293 cells cultured in fully synthetic media. No animal products are used during the production process.

Chikungunya Virus VLP (E1 E2 C protein)

CHIKV-VLP-500 0.5
EUR 2636.98
Description: Recombinant Chikungunya Virus-Like Particles produced in 293 human cells. Chikungunya structural proteins are transiently expressed to form a particle comprising E1, E2 and capsid protein, without the non-structural proteins or genome. Resultant particles are totally non-infectious. This product is prepared from HEK293 cells cultured in fully synthetic media. No animal products are used during the production process.

Chikungunya virus E1 protein (A226V mutant)

30-1943 100 ug
EUR 1500
Description: Recombinant Chikungunya virus E1 protein - A266V mutant

Chikungunya Virus Human IgG Assay Control

BC148G 1 mL
EUR 105

Chikungunya Virus Human IgM Assay Control

BC148M 1 mL
EUR 105

ELISA kit for Human Chikungunya Virus IgG (CHIKV IgG)  Kit

KTE62972-48T 48T
EUR 424.8
Description: Quantitative sandwich ELISA for measuring Human Chikungunya Virus IgG (CHIKV IgG)  Kit in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.

ELISA kit for Human Chikungunya Virus IgG (CHIKV IgG)  Kit

KTE62972-5platesof96wells 5 plates of 96 wells
EUR 2702.4
Description: Quantitative sandwich ELISA for measuring Human Chikungunya Virus IgG (CHIKV IgG)  Kit in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.

ELISA kit for Human Chikungunya Virus IgG (CHIKV IgG)  Kit

KTE62972-96T 96T
EUR 686.4
Description: Quantitative sandwich ELISA for measuring Human Chikungunya Virus IgG (CHIKV IgG)  Kit in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.

ELISA kit for Human Chikungunya Virus IgM (CHIKV IgM)  Kit

KTE62973-48T 48T
EUR 424.8
Description: Quantitative sandwich ELISA for measuring Human Chikungunya Virus IgM (CHIKV IgM)  Kit in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.

ELISA kit for Human Chikungunya Virus IgM (CHIKV IgM)  Kit

KTE62973-5platesof96wells 5 plates of 96 wells
EUR 2702.4
Description: Quantitative sandwich ELISA for measuring Human Chikungunya Virus IgM (CHIKV IgM)  Kit in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.

ELISA kit for Human Chikungunya Virus IgM (CHIKV IgM)  Kit

KTE62973-96T 96T
EUR 686.4
Description: Quantitative sandwich ELISA for measuring Human Chikungunya Virus IgM (CHIKV IgM)  Kit in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.

Chikungunya virus gpE1 protein (A226V mutant)

30-1941 100 ug
EUR 1560
Description: Recombinant Chikungunya virus Glycoprotein E1 protein - A226V mutant

Polyclonal Antibody to Chikungunya Virus (CHIKV)

PAX256Ge08 100ul
EUR 385

Monoclonal Antibody to Chikungunya Virus (CHIKV)

MAX256Ge21 100ul
EUR 455

Chikungunya Virus E1 Envelope Protein (HEK293)

CHIKV-E1-100 0.1
EUR 401.87
Description: Recombinant Chikungunya virus envelope protein 1 (E1), expressed in HEK293 mammalian cells.

Chikungunya Virus E1 Envelope Protein (HEK293)

CHIKV-E1-500 0.5
EUR 1542.39
Description: Recombinant Chikungunya virus envelope protein 1 (E1), expressed in HEK293 mammalian cells.

Hi-PCR® Chikungunya SYBr PCR Kit

MBPCR083-10R 1 unit
EUR 48.94
Description: Hi-PCR® Chikungunya SYBr PCR Kit

Hi-PCR® Chikungunya SYBr PCR Kit

MBPCR083-50R 1 unit
EUR 208.63
Description: Hi-PCR® Chikungunya SYBr PCR Kit

Chikungunya virus Non-structural polyprotein,

1-CSB-EP810351CJAT
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  • 100ug
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  • 1MG
  • 200ug
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  • 50ug
Description: Recombinant Chikungunya virus Non-structural polyprotein,Partial expressed in E.coli

Chikungunya Virus E1 Envelope Protein (Insect Cells)

REC31767-100 0.1
EUR 676.15
Description: Recombinant Chikungunya virus CHIKV) wild-type E1 protein (Asian genotype) is produced in insect cells. It is composed of amino acids 67-482 (Gene Accession: EU441882) of the Chikungunya E1 protein and the secreted and soluble recombinant protein (MW ~50

Chikungunya Virus (CHIKV) Polyclonal Antibody (Pan-species)

4-PAX256Ge08
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  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
Description: A Rabbit polyclonal antibody against Pan-species Chikungunya Virus (CHIKV)

Chikungunya Virus E1 Envelope Protein (CHIKV-E1) Protein

abx120021-100l 100 µl Ask for price

Chikungunya Virus E1 Envelope Protein (CHIKV-E1) Protein

abx120021-20l 20 µl
EUR 1387.5

Chikungunya Virus E1 Envelope Protein (CHIKV-E1) Protein

abx120021-50l 50 µl Ask for price

Chikungunya virus human monoclonal antibody, clone E26D9

DDX9100HD01 1 mg Ask for price

Chikungunya virus human monoclonal antibody, clone E26D9

DDX9100P-100 100 µg Ask for price

Chikungunya virus human monoclonal antibody, clone E26C4

DDX9102HD01 1 mg Ask for price

Chikungunya virus human monoclonal antibody, clone E26C4

DDX9102P-100 100 µg Ask for price

OPNB00001-100UG - Chikungunya virus E1 Envelope protein

OPNB00001-100UG 100ug
EUR 504

OPNB00001-500UG - Chikungunya virus E1 Envelope protein

OPNB00001-500UG 500ug
EUR 1987

Chikungunya Virus (CHIKV) Polyclonal Antibody (Pan-species), PE

4-PAX256Ge08-PE
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  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
Description: A Rabbit polyclonal antibody against Pan-species Chikungunya Virus (CHIKV). This antibody is labeled with PE.

Chikungunya Virus (CHIKV) Polyclonal Antibody (Pan-species), APC

4-PAX256Ge08-APC
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  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
Description: A Rabbit polyclonal antibody against Pan-species Chikungunya Virus (CHIKV). This antibody is labeled with APC.

Chikungunya Virus (CHIKV) Polyclonal Antibody (Pan-species), Cy3

4-PAX256Ge08-Cy3
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  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
Description: A Rabbit polyclonal antibody against Pan-species Chikungunya Virus (CHIKV). This antibody is labeled with Cy3.

Chikungunya Virus (CHIKV) Polyclonal Antibody (Pan-species), HRP

4-PAX256Ge08-HRP
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  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
Description: A Rabbit polyclonal antibody against Pan-species Chikungunya Virus (CHIKV). This antibody is labeled with HRP.

GWB-7CF864-96W - Chikungunya Virus IgM u-capture ELISA

GWB-7CF864-96W 96Wells
EUR 559

Chikungunya Virus (CHIKV) Polyclonal Antibody (Pan-species), FITC

4-PAX256Ge08-FITC
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  • 100ul
  • 10ml
  • 1ml
  • 200ul
  • 20ul
Description: A Rabbit polyclonal antibody against Pan-species Chikungunya Virus (CHIKV). This antibody is labeled with FITC.
On this examine, particular person exosomes had been noticed on a trajectory when passing by the laser beam sheet and had been counted for 10 min at a relentless circulate velocity. The entire evaluation time was lower than 1.5 h together with the steps required to take away massive particles and permit response with the antibody. The outcomes for 2 samples ready with and with out the isolation of exosomes confirmed a lack of exosomes within the isolation step. We additionally decided the variety of the exosomes secreted by the cells to a tradition medium throughout cultivation. As anticipated, the overall variety of exosomes in a tradition medium elevated with a rise within the cultivation time, and the variety of exosomes launched each 12 h both remained fixed or confirmed not more than a slight improve for so long as 72 h. It was unclear whether or not the quantity exosomes was depending on the cell inhabitants at confluences of 10-60%.
Gina

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