Optimization of microRNA Acquirement from Seminal Plasma and Identification of Diminished Seminal microRNA-34b as Indicator of Low Semen Concentration

Approximately 10-15% of couples who want to conceive suffer from subfertility, while in 30% of these cases, male factors play a role. The level of specific microRNAs in seminal plasma, including those involved in spermatogenesis, can serve as a parameter indicative of subfertility. We first optimized protocols to obtain microRNAs from seminal plasma.

Next, using a test strategy is validated in a group of men, we aim to identify microRNAs whose levels associated with semen motility and concentration. By qPCR, 742 microRNAs are profiled in normozoospermic three samples, three samples of sperm and low sperm motility (asthenozoospermia), and three with semen low concentration (oligozoospermia). MicroRNAs show a significant difference between the groups further validated in a second group consisting of 40 samples with normozoospermia (control group), 47 samples with asthenozoospermia, and 19 samples with oligozoospermia (which is 74% too low motility).

MicroRNA highest results obtained with biofluids RNA extraction kits, with the inclusion of MS2 RNA and proteinase K treatment operator for the protocol, and when 50 uL seminal plasma is used as input. Exosome RNA isolation before extraction does not lead to improved results. In the test cohort, 236 microRNAs can be detected, of which 54 microRNAs showed differences between groups. Five microRNAs analyzed in the validation group. the level of MiR-34b-5p in the control group was significantly higher compared with asthenozoospermia group (p <0.05) and compared with oligozoospermia group (p <0.001).

We optimized the acquisition of seminal plasma microRNA and microRNA levels were identified in relation to sperm concentration and motility. As the latest human and rat studies showed that miR-34 family is a marker of low concentration of semen and is very important in spermatogenesis, seminal plasma miR-34b-5p may be a suitable candidate for further study as a marker of male subfertility.

 Optimization of microRNA Acquirement from Seminal Plasma and Identification of Diminished Seminal microRNA-34b as Indicator of Low Semen Concentration
Optimization of microRNA Acquirement from Seminal Plasma and Identification of Diminished Seminal microRNA-34b as Indicator of Low Semen Concentration

When Less Is More: Special Capture and Analysis in the Plasma Tumor Exosom Liquid Biopsy Improves Sensitivity for Detection of Multiple androgen receptor Comprehensive Phenotypes in Advanced Prostate Cancer Patients

We evaluate the advantages and reliability of new protocols for enriching tumor extracellular vesicles (EV), allowing blood-based test for noninvasive parallel profiles of some of the androgen receptor (AR) gene changes. Three AR clinically relevant variants associated with response / resistance to standard-of-care treatment (AR-V7 transcripts, a point mutation AR T878A and AR gene amplification) were evaluated by digital PCR in 15 samples from patients affected by Castration-Resistant Prostate Cancer (CRPC).

Plasma is processed to obtain circulating RNA and DNA using protocols based enrichment EVS tumor through immuno-affinity and peptide-affinity compared with the generic extraction kit. Our results indicate that the immuno-affinity enrichment before RNA extraction clearly exceed the generic isolation method in the detection of the AR-V7, also allow for the differences between the respondents (R) and non-responders (NR) patients.The T878A mutation is detected, as a whole, in nine out of 15 samples, and no one approach alone is capable of expressing the mutation in all samples store, indicating that the method employed complementary.

Exosome precipitation from Plasma + Thrombin

EPStep-T 5 ml
EUR 403.2

ExoQuick Exosome RNA Column Purification kit

EQ808A-1 20 preps
EUR 482.4

SeraMir Exosome RNA Purification kit (5ml ExoQuick and 20 exoRNA columns)

RA806A-1 20 preps
EUR 664.8

Exosome precipitation from Plasma and Serum

EPStep-PS 5 ml
EUR 356.4

Thrombin Plasma prep for Exosome precipitation (500 ul at 500U/mL)

TMEXO-1 100 reactions
EUR 699.6

ExoQuick-LP for lipoprotein pre-clear & exosome isolation

EXOLP5A-1 5 reactions
EUR 639.6

TumorExoRNA? Tumor-derived exosome immunocapture and RNA extraction kit (20 reactions)

K1221-20 each
EUR 1462.8

ExoPure? 0.4 micron Immunobeads (Overall Exosome Isolation, cell media, 20 reactions)

M1030-20 each
EUR 1546.8

ExoPure? 1.0 micron Immunobeads (Overall Exosome Isolation, cell media, 20 reactions)

M1031-20 each
EUR 1546.8

TumorExoPure? 0.4 micron Immunobeads (Tumor-derived Exosome Isolation, biofluids, 20 reactions)

M1032-20 each
EUR 1618.8

TumorExoPure? 1.0 micron Immunobeads (Tumor-derived Exosome Isolation, biofluids, 20 reactions)

M1033-20 each
EUR 1618.8

MouseExoPure? 0.4 micron Immunobeads (Overall Exosome Isolation, mouse biofluids, 20 reactions)

M1034-20 each
EUR 1495.2

MouseExoPure? 1.0 micron Immunobeads (Overall Exosome Isolation, mouse biofluids, 20 reactions)

M1035-20 each
EUR 1495.2

Formalin Solution (20%)

FRL125 125 ml
EUR 75.6

Formalin Solution (20%)

FRL500 500 ml
EUR 98.4

Formalin Solution (20%)

FRL999 1000 ml
EUR 121.2

20% SDS Solution

S0793-050 500ml
EUR 144

20% SDS Solution

S0793-100 2X500ml
EUR 199.2

ExoQuick PLUS Exosome Purification Kit - for serum and plasma

EQPL10A-1 10 reactions
EUR 694.8

MouseExoPure? 0.4 micron Immunobeads (Overall Exosome Isolation, mouse cell media, 20 reactions)

M1036-20 each
EUR 1546.8

MouseExoPure? 1.0 micron Immunobeads (Overall Exosome Isolation, mouse cell media, 20 reactions)

M1037-20 each
EUR 1546.8

ExoPure? 1.0 micron Immunobeads (Overall Exosome Isolation, plasma, urine, serum, 20 reactions)

M1039-20 each
EUR 1546.8

Proteinase K, Recombinant, 20 mg/ml Solution, Molecular Grade

9211-100 each
EUR 1495.2

Proteinase K, Recombinant, 20 mg/ml Solution, Molecular Grade

9211-25 each
EUR 510

Proteinase K, Recombinant, 20 mg/ml Solution, Molecular Grade

9211-5 each
EUR 196.8

ExoDNAPS? Circulating and Exosome-associated DNA Extraction Kit (Human Plasma/Serum, 20 reactions)

K1230-20 each
EUR 861.6

ExoDNAUC? Circulating and Exosome-associated DNA Extraction Kit (Urine/Cell Media, 20 reactions)

K1231-20 each
EUR 861.6

ExoQuick-TC PLUS Exosome Purification Kit - for tissue culture media

EQPL10TC-1 10 reactions
EUR 694.8

HSA Solution 20% (cGMP)

AK8214-0100 100mL Ask for price

Arabinose induction solution (20%)

MN850A-1 20 ml
EUR 152.4

Glycerol, 20% Sterile Solution

GR103065 100 mL
EUR 70.8

100ML SDS Solution 20%

NAT1228 100ML
EUR 36.82

450ML SDS Solution 20%

NAT1230 450ML
EUR 90.06

SDS, 20% Liquid Solution

18-234 500ml/Unit
EUR 334

SDS, 20% Liquid Solution

18-234B 1000ml/Unit
EUR 390

Puromycin DiHCl Solution (10 mg/mL in 20 mM HEPES)

P025-P026-P025-5x1ML P025-5 x 1 mL
EUR 148.8

Puromycin DiHCl Solution (10 mg/mL in 20 mM HEPES)

P026-P026-20ML P026-20 mL
EUR 337.2

TotalExoRNA? Overall exosome immunocapture and RNA extraction kit (Biological Fluids/Cell Media, 20 reactions)

K1220-20 each
EUR 1462.8

Dispase Solution, 1mg/ml

CA092-010 100ml
EUR 134.4

Transferrin Solution, 10mg/ml

CA205-005 5x1ml
EUR 138

cleaning solution 500 ml

B530 ea
EUR 36

Kanamycin solution (50mg/ml)

MN800A-1 20 ml
EUR 152.4

Gentamicin Solution, Gentamicin Solution, 50 mg/mL

CCM1123-010 10 mL
EUR 76.86

TBE 10X Liquid Solution Ultra Pure Grade Solution

20-196 4 Liters/Unit
EUR 400

ExoQuick Exosome Isolation and RNA Purification kit (for Serum and Plasma)

EQ806A-1 20 preps
EUR 664.8

ExoQuick Exosome Isolation and RNA Purification kit (for Tissue Culture Media)

EQ806TC-1 10 preps
EUR 664.8

ExoQuick TC 10ml

EXOTC10A-1 10 ml
EUR 478.8

ExoQuick TC 50ml

EXOTC50A-1 50 ml
EUR 1466.4

99445-20 DCT 20 X 150MM

99445-20 250/pk
EUR 142.8
Description: Disposable Culture Tubes; DCT's, CGW

20 x Synovial Fluid Solution

GR103059 100 mL
EUR 84

SDS, 20%(w/v) solution

SD8119 100ml
EUR 76.7

Albumin, Bovine 20mg/ml solution

CH006 1.0 ml
EUR 170.4

Albumin, Bovine 20mg/ml solution

CH007 5.0 ml
EUR 254.4

Chloramphenicol Solution (50 mg/ml)

C090-10ML 10 mL
EUR 70.8

Chloramphenicol Solution (50 mg/ml)

C090-10x10ML 10 x 10 mL
EUR 349.2

Sodium Selenite Solution,100mg/ml

CA056-010 100ml
EUR 121.2

DNase I Solution, 1mg/ml

CA095-005 5x1ml
EUR 165.6

BSA Blocking Solution, 2 mL

BLK-002 2 mL
EUR 106.21
Description: The BSA solution is blocking gold nanoparticles following conjugation reactions.

BSA Blocking Solution, 10 mL

BLK-010 10 mL
EUR 202.3
Description: The BSA solution is blocking gold nanoparticles following conjugation reactions.

pepsine cleaning solution 250 ml

B231 ea
EUR 54

pepsine cleaning solution 1000 ml

B232 ea
EUR 118.8

RNase A Solution (10mg/ml)

RB0474 1ml
EUR 66.26

ExoDNAPS? Conc Exosome DNA Extraction Kit

K1243-20 each
EUR 992.4

ExoDNAUC? Conc Exosome DNA Extraction Kit

K1244-20 each
EUR 992.4

LiqBiopsy Tumor Exosome DNA Kit I

K1255-20 each
EUR 1201.2

LiqBiopsy Tumor Exosome DNA Kit II

K1256-20 each
EUR 1266

DNA Precipitation Buffer

D106 55 ml
EUR 210
Description: DNA Precipitation Buffer by Cygnus Technologies is available in Europe via Gentaur.

TMB for Precipitation

42R-TB100x 1000 ml
EUR 645.6
Description: High sensitivity TMB Substrate for Precipitation assays

EpiNext Hi-Fi cDNA Synthesis Kit  

P-9004
  • EUR 381.90
  • EUR 227.70
  • 20 Samples
  • 20 Reactions

20(S)-NotoginsenosideR2

N2028-20 20 mg
EUR 756
Description: Extracted from Pseudo-ginseng;Store the product in sealed, cool and dry condition

ExoPure? Reagent (Overall Exosome Isolation, biological fluids)

M1001-20 each
EUR 1260

Human Serum Albumin protein (20% Solution)

30-1138 50 mL
EUR 632.4
Description: Human Serum Albumin, 20% solution

Potassium Iodide 20% Solution w/v

KI20WV1 1L
EUR 72.96

Potassium Thiocyanate Solution 20% w/v

KT20WV1 1L
EUR 74.1

Barium Chloride Solution 20% w/v

WTR081 1L
EUR 67.26

50 ML KANAMYCIN SULFATE, LIQUID, 5,000 µG/ML SOLUTION

30-006-CF 50 mL/pk
EUR 117.6
Description: Media Catalog; Cell Culture Reagents

Ethidium Bromide, 10mg/ml Stock Solution

E0205-001 10ml
EUR 105.6

Ethidium Bromide, 10mg/ml Stock Solution

E0205-002 20ml
EUR 153.6

Fibronectin, Bovine, Stabilized Solution , 1mg/ml

CA082-001 1ml
EUR 375.6

Fibronectin, Human, Stabilized Solution, 1mg/ml

CA083-001 1ml
EUR 392.4

Vitronectin , Human , Stabilized Solution, 0.5mg/ml

CA085-050 50ul
EUR 250.8

Papain Solution In PBS, 1mg/ml

CA093-002 20ml Ask for price

Insulin Solution, 1mg/ml , Animal Free

CA204-005 5x1ml
EUR 138

500 ML L-GLUTAMINE 200MM SOLUTION

25-005-CV 500 mL/pk
EUR 116.4
Description: Media Catalog; Cell Culture Reagents

100 ML SODIUM BICARBONATE, 7.5% SOLUTION

25-035-CI 100 mL/pk
EUR 69.6
Description: Media Catalog; Cell Culture Reagents

buffer solution 500 ml 4.00 ph

B504 ea
EUR 36

buffer solution 500 ml 7.00 ph

B507 ea
EUR 36

buffer solution 500 ml 10.01 ph

B510 ea
EUR 36

buffer solution 500 ml 12.00 ph

B512 ea
EUR 36

calibration solution 500 ml 200ms/cm

B566B ea
EUR 54

buffer solution 50 ml 4.00 ph

B004 ea
EUR 25.2

buffer solution 50 ml 7.00 ph

B007 ea
EUR 25.2

buffer solution 50 ml 10.00 ph

B010 ea
EUR 25.2

calibration solution 50 ml 200ms/cm

B066 ea
EUR 28.8

Linear Poly Acrylamide Solution (5mg/ml)

LK548 1ml
EUR 75.66

Hemocyanin-Keyhole Limpet (KLH) subunits, solution

6287-20 each
EUR 418.8

20(R)Ginsenoside?Rh1

N2192-20 20 mg
EUR 616.8
Description: Extracted from Panax ginseng C. A. Mey. dried roots;Store the product in sealed, cool and dry condition

20(R)Ginsenoside Rg2

N2195-20 20 mg
EUR 268.8
Description: Extracted from Panax ginseng C. A. Mey. dried roots;Store the product in sealed, cool and dry condition

20(R)Ginsenoside Rg3

N2198-20 20 mg
EUR 616.8
Description: Extracted from Panax ginseng C.A.Mey. roots;Store the product in sealed, cool and dry condition

FGF-20, human recombinant

4589-20 each
EUR 332.4

D(+)Glucose 20 % 1000 ml

12-9165-5 1000 ml
EUR 170.4

RNaseA50 mg/ml -20 C

FYG106-130UL 130 µl Ask for price

RNaseA50 mg/ml -20 C

FYG106-200UL 200 µl Ask for price

RNaseA50 mg/ml -20 C

FYG106-50UL 50 µl Ask for price

Dubos Med Albu 20 Ml

215334 PK10
EUR 134.52

SeraMir Exosome RNA Purification kit (10ml ExoQuick-TC and 10 exoRNA columns) 10 preps

RA806TC-1 10 preps
EUR 664.8

Reagecon coloured buffer solution - pH at 20 C - pH 4.00 +/- 0.01 - 500 mL bottle

DD440460 EACH
EUR 33.06

Reagecon coloured buffer solution - pH at 20 C - pH 10.00 +/- 0.01 - 500 mL bottle

DD440464 EACH
EUR 33.06

EtBr Destaining Bag Kit Activation Solution Included

20-277 25 Bags/Unit
EUR 394

Chicken IgY precipitation reagent

85R-1010 4 ml
EUR 664.8
Description: Chicken IgY precipitation reagent for use in immunoassays

PEG Virus Precipitation Kit

K2190-200 200 preparations
EUR 1270.8

PEG Virus Precipitation Kit

K2190-50 50 preparations
EUR 486

50ML Protein Precipitation Kit

NAT1254 50ML
EUR 237.12

PEG Virus Precipitation Kit

K904-200 each
EUR 1201.2

PEG Virus Precipitation Kit

K904-50 each
EUR 451.2

Pluronic f-127 20% solution in DMSO

59004 1ML
EUR 79.2
Description: Minimum order quantity: 1 unit of 1ML

Hoechst 33258 *20 mM solution in water*

17525 5 mL
EUR 158.4

Hoechst 33342 *20 mM solution in water*

17535 5 mL
EUR 158.4

Hoechst 34580 *20 mM solution in water*

17538 100 uL
EUR 262.8

Sodium dodecyl sulfate, 20% solution in water

GD0990-100ML 100 ml
EUR 64.8

Sodium dodecyl sulfate, 20% solution in water

GD0990-1L 1 l
EUR 151.2

Sodium dodecyl sulfate, 20% solution in water

GD0990-250ML 250 ml
EUR 84

Sodium dodecyl sulfate, 20% solution in water

GD0990-2L 2 l
EUR 246

Tween® 20, MegaPure? Detergent, 10% Solution

2125-100 each
EUR 242.4

Tween® 20, MegaPure? Detergent, 10% Solution

2125-50 each
EUR 176.4

Tween® 20, MegaPure? Detergent, 10% Solution

2125-500 each
EUR 836.4

Tween® 20, MegaPure? Detergent, 10% Solution

2125-set each
EUR 222

20 x ELISA Wash Buffer

GR103014-20 1 L
EUR 189.6

Hanks Balanced Salt Solution 500ml

20-021-CVR PK6
EUR 221.16

ExoPure? 0.4 micron Immunobeads (Overall Exosome Isolation, plasma, urine, serum)

M1038-20 each
EUR 1546.8

Puromycin Solution, 10mg/ml , Cell Cultere Tested

CR026-001 10ml
EUR 241.2

Fetuin Solution, 1mg/ml in DMEM-F12

CA065-005 5x1ml
EUR 116.4

CometAssay Lysis Solution (2 x 500 mL)

4250-050-01 1 L
EUR 219.6

PBS solution pH 7.4, 1000 ml/bottle

10-9402-10 1000 ml/bottle
EUR 193.2

dsGreen Gel Staining Solution, 10000×, 0.5 mL

10010 0.5 mL
EUR 106.8

calibration solution 500 ml 0.01 m kcl

B560 ea
EUR 36

calibration solution 500 ml 0.1 m kcl

B561 ea
EUR 36

calibration solution 500 ml 1 m kcl

B562 ea
EUR 36

calibration solution 50 ml 0.01 m kcl

B060 ea
EUR 25.2

calibration solution 50 ml 0.1 m kcl

B061 ea
EUR 25.2

calibration solution 50 ml 1 m kcl

B062 ea
EUR 25.2

redox standard solution 50 ml 124 mv

B071 ea
EUR 26.4

redox standard solution 50 ml 358 mv

B072 ea
EUR 26.4

redox standard solution 500 ml 124 mv

B571 ea
EUR 43.2

redox standard solution 500 ml 358 mv

B572 ea
EUR 43.2

Spectrophotometric Standard Protein Solution, 3 mg/ml

GR103032 5 x 1 mL
EUR 67.2

X-Gal Solution, 40 mg/ml concentrate

X0321-005 5X1ml
EUR 138

X-Gal Solution, 40 mg/ml concentrate

X0321-020 20ml
EUR 162

TAE 50X Liquid Concentrate Ultra Pure Grade Solution

20-193 1.6 Liters/Unit
EUR 405

TAE 50X Liquid Concentrate Ultra Pure Grade Solution

20-194 4 Liters/Unit
EUR 566

TAE 25X Liquid Concentrate Ultra Pure Grade Solution

20-195 1.6 Liters/Unit
EUR 379

Tris-Glycine 10X Liquid Solution Ultra Pure Grade

20-246 4 Liters/Unit
EUR 402

TG-SDS 10X Liquid Solution Ultra Pure Grade

20-247 4 Liters/Unit
EUR 403

Complete Freund's Adjuvant, 20 mg/ml

7024 20 mg/ml x 5 ml
EUR 472.38
Description: Complete Freund's Adjuvant

20UL FILTER TIPS FOR P-20

TF-20 1000/pk
EUR 501.6
Description: Filter Tip; Filter Tips Pipette - Axygen

Anti-Cat Whole Serum | RWS-20

RWS-20 1.0 mL
EUR 153
Description:

Anti-Cat Whole Serum | RWS-20 | Immunology Consultants Laboratory

Host: Rabbit

Format: Whole Serum

Product Type: Primary Antibody

Antibody Clonality: Polyclonal

AR amplification CRPC were detected in most samples were analyzed using either a cell-free DNA (cfDNA) or exosome isolation kit (80%). We show that the selective isolation of exosomes enriched subset for the origin of the tumor, rather than the total analysis of exosomes plasma or plasma total nucleic acids, increased sensitivity and specificity to detect specific changes in circulation.