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 binding buffer for ELISAs (20 ml)

EXOEL-BB-20 20 ml
EUR 219.00
  • Category: Exosomes

PEG-it Virus Precipitation Solution (100 mL aliquot)

LV810A-1 100 ml
EUR 382.00
  • Category: Lentiviral Technology

PEG-it Virus Precipitation Solution (250 mL aliquot)

LV825A-1 250 ml
EUR 777.00
  • Category: Lentiviral Technology

ExoQuick-CG cGMP-grade Exosome Isolation reagent (50 ml)

EXOCG50A-1 50 ml
EUR 1181.00
  • Category: Exosomes

Exosome precipitation from Plasma + Thrombin

EPStep-T 5 ml
EUR 336.00

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

RA806A-1 20 preps
EUR 554.00
  • Category: MicroRNA Tools

ExoQuick Exosome RNA Column Purification kit

EQ808A-1 20 preps
EUR 402.00

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

TMEXO-1 100 reactions
EUR 583.00
  • Category: Exosomes

Exosome precipitation from Plasma and Serum

EPStep-PS 5 ml
EUR 297.00

ExoQuick-LP for lipoprotein pre-clear & exosome isolation

EXOLP5A-1 5 reactions
EUR 533.00
  • Category: Exosomes

Formalin Solution (20%)

FRL125 125 ml
EUR 63.00

Formalin Solution (20%)

FRL500 500 ml
EUR 82.00

Formalin Solution (20%)

FRL999 1000 ml
EUR 101.00

20% SDS Solution

S0793-050 500ml
EUR 120.00

20% SDS Solution

S0793-100 2X500ml
EUR 166.00

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

9211-100
EUR 1246.00

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

9211-25
EUR 425.00

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

9211-5
EUR 164.00

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

K1221-20
EUR 1219.00

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

M1030-20
EUR 1289.00

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

M1033-20
EUR 1349.00

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

M1034-20
EUR 1246.00

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

M1031-20
EUR 1289.00

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

M1032-20
EUR 1349.00

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

M1035-20
EUR 1246.00

ExoQuick PLUS Exosome Purification Kit - for serum and plasma

EQPL10A-1 10 reactions
EUR 579.00
  • Category: Exosomes

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

P025-P026-P025-5x1ML P025-5 x 1 mL
EUR 124.00

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

P026-P026-20ML P026-20 mL
EUR 281.00

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

M1036-20
EUR 1289.00

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

M1037-20
EUR 1289.00

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

M1039-20
EUR 1289.00

cleaning solution 500 ml

B530 ea
EUR 30.00

Dispase Solution, 1mg/ml

CA092-010 100ml
EUR 112.00

Transferrin Solution, 10mg/ml

CA205-005 5x1ml
EUR 115.00

Kanamycin solution (50mg/ml)

MN800A-1 20 ml
EUR 127.00
  • Category: Minicircle Technology

HSA Solution 20% (cGMP)

AK8214-0100 100mL Ask for price

100ML SDS Solution 20%

NAT1228 100ML
EUR 77.00

450ML SDS Solution 20%

NAT1230 450ML
EUR 115.00

Arabinose induction solution (20%)

MN850A-1 20 ml
EUR 127.00
  • Category: Minicircle Technology

Gentamicin Solution, Gentamicin Solution, 50 mg/mL

CCM1123-010 10 mL
EUR 64.05
  • Product category: Culture Media

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

K1230-20
EUR 718.00

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

K1231-20
EUR 718.00

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

EQPL10TC-1 10 reactions
EUR 579.00
  • Category: Exosomes

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

K1220-20
EUR 1219.00

pepsine cleaning solution 250 ml

B231 ea
EUR 45.00

pepsine cleaning solution 1000 ml

B232 ea
EUR 99.00

Sodium Selenite Solution,100mg/ml

CA056-010 100ml
EUR 101.00

DNase I Solution, 1mg/ml

CA095-005 5x1ml
EUR 138.00

Chloramphenicol Solution (50 mg/ml)

C090-10ML 10 mL
EUR 59.00

Chloramphenicol Solution (50 mg/ml)

C090-10x10ML 10 x 10 mL
EUR 291.00

BSA Blocking Solution, 2 mL

BLK-002 2 mL
EUR 88.51
  • Product Category: Gold Ribbon, Gold Conjugation
Description: The BSA solution is blocking gold nanoparticles following conjugation reactions.

BSA Blocking Solution, 10 mL

BLK-010 10 mL
EUR 168.58
  • Product Category: Gold Ribbon, Gold Conjugation
Description: The BSA solution is blocking gold nanoparticles following conjugation reactions.

Albumin, Bovine 20mg/ml solution

CH006 1.0 ml
EUR 142.00

100 ML 45% GLUCOSE SOLUTION

25-037-CI 100 mL/pk
EUR 46.00
Description: Media Catalog; Cell Culture Reagents

Lentivirus Concentration Solution (100 mL)

P904C NULL
EUR NULL

Retrovirus Concentration Solution (100 mL)

P905C NULL
EUR NULL

RNase A Solution (10mg/ml)

RB0474 1ml
EUR 55.22
  • Product category: Proteins/Enzymes/Endonucleases

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

EQ806A-1 20 preps
EUR 554.00

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

EQ806TC-1 10 preps
EUR 554.00

SDS, 20%(w/v) solution

SD8119 100ml
EUR 63.92
  • Product category: Biochemicals/Detergents/Surfactants

99445-20 DCT 20 X 150MM

99445-20 250/pk
EUR 119.00
Description: Disposable Culture Tubes; DCT's, CGW

ExoQuick TC 10ml

EXOTC10A-1 10 ml
EUR 399.00
  • Category: Exosomes

ExoQuick TC 50ml

EXOTC50A-1 50 ml
EUR 1222.00
  • Category: Exosomes

10 ML GENTAMICIN SULFATE, LIQUID 50 MG/ML SOLUTION

30-005-CR 10 mL/pk
EUR 433.00
Description: Media Catalog; Cell Culture Reagents

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

30-006-CF 50 mL/pk
EUR 98.00
Description: Media Catalog; Cell Culture Reagents

Exosome isolation Solution - Serum/ Plasma (easy version of P101, 95% pure exosome)

P101EZ NULL
EUR NULL

buffer solution 50 ml 4.00 ph

B004 ea
EUR 21.00

buffer solution 50 ml 7.00 ph

B007 ea
EUR 21.00

buffer solution 50 ml 10.00 ph

B010 ea
EUR 21.00

calibration solution 50 ml 200ms/cm

B066 ea
EUR 24.00

buffer solution 500 ml 2.00 ph

B502 ea
EUR 30.00

buffer solution 500 ml 4.00 ph

B504 ea
EUR 30.00

buffer solution 500 ml 7.00 ph

B507 ea
EUR 30.00

buffer solution 500 ml 10.01 ph

B510 ea
EUR 30.00

buffer solution 500 ml 12.00 ph

B512 ea
EUR 30.00

Fibronectin, Bovine, Stabilized Solution , 1mg/ml

CA082-001 1ml
EUR 313.00

Fibronectin, Human, Stabilized Solution, 1mg/ml

CA083-001 1ml
EUR 327.00

Vitronectin , Human , Stabilized Solution, 0.5mg/ml

CA085-050 50ul
EUR 209.00

Papain Solution In PBS, 1mg/ml

CA093-002 20ml Ask for price

Insulin Solution, 1mg/ml , Animal Free

CA204-005 5x1ml
EUR 115.00

calibration solution 500 ml 200ms/cm

B566B ea
EUR 45.00

500 ML L-GLUTAMINE 200MM SOLUTION

25-005-CV 500 mL/pk
EUR 97.00
Description: Media Catalog; Cell Culture Reagents

100 ML MEM VITAMINS 100X SOLUTION

25-020-CI 100 mL/pk
EUR 67.00
Description: Media Catalog; Cell Culture Reagents

100 ML SODIUM BICARBONATE, 7.5% SOLUTION

25-035-CI 100 mL/pk
EUR 58.00
Description: Media Catalog; Cell Culture Reagents

7-AAD solution, 1 mg/mL

40084 1mL
EUR 212.00
Description: Minimum order quantity: 1 unit of 1mL

Ethidium Bromide, 10mg/ml Stock Solution

E0205-001 10ml
EUR 88.00

Ethidium Bromide, 10mg/ml Stock Solution

E0205-002 20ml
EUR 128.00

Linear Poly Acrylamide Solution (5mg/ml)

LK548 1ml
EUR 63.05
  • Product category: PCR Related/Nucleic Acid Co-precipitants

500 ML HBSS (HANK'S BALANCED SALT SOLUTION), 10X WITHOUT SODIUM BICARBONATE, CALCIUM AND MAGNESIUM

20-021-CVR 500 mL/pk
EUR 105.00
Description: Media Catalog; Buffered Salt-Salution

Human Serum Albumin protein (20% Solution)

30-1138 50 mL
EUR 527.00
Description: Human Serum Albumin, 20% solution

D(+)Glucose 20 % 1000 ml

12-9165-5 1000 ml
EUR 142.00

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

TMB for Precipitation

42R-TB100x 1000 ml
EUR 538.00
Description: High sensitivity TMB Substrate for Precipitation assays

DNA Precipitation Buffer

D106 55 ml
EUR 175.00
  • Product category: DNA and RNA extraction and purification
Description: DNA Precipitation Buffer by Cygnus Technologies is available in Europe via Gentaur.

ExoDNAPS? Conc Exosome DNA Extraction Kit

K1243-20
EUR 827.00

ExoDNAUC? Conc Exosome DNA Extraction Kit

K1244-20
EUR 827.00

LiqBiopsy Tumor Exosome DNA Kit I

K1255-20
EUR 1001.00

LiqBiopsy Tumor Exosome DNA Kit II

K1256-20
EUR 1055.00

20(S)-NotoginsenosideR2

N2028-20 20 mg
EUR 630.00
Description: Extracted from Pseudo-ginseng;Store the product in sealed, cool and dry condition

Exosome isolation Solution - Cell Media/ Urine (easy version of P100, 95% pure exosome)

P100EZ NULL
EUR NULL

Exosome isolation Solution - Serum/ Plasma (easy version of P101, 95% pure exosome) sample

P101EZ--sample NULL
EUR NULL

ExoPure? Reagent (Overall Exosome Isolation, biological fluids)

M1001-20
EUR 1050.00

Hemocyanin-Keyhole Limpet (KLH) subunits, solution

6287-20
EUR 349.00

calibration solution 50 ml 0.01 m kcl

B060 ea
EUR 21.00

calibration solution 50 ml 0.1 m kcl

B061 ea
EUR 21.00

calibration solution 50 ml 1 m kcl

B062 ea
EUR 21.00

redox standard solution 50 ml 124 mv

B071 ea
EUR 22.00

redox standard solution 50 ml 358 mv

B072 ea
EUR 22.00

Fetuin Solution, 1mg/ml in DMEM-F12

CA065-005 5x1ml
EUR 97.00

calibration solution 500 ml 0.01 m kcl

B560 ea
EUR 30.00

calibration solution 500 ml 0.1 m kcl

B561 ea
EUR 30.00

calibration solution 500 ml 1 m kcl

B562 ea
EUR 30.00

redox standard solution 500 ml 124 mv

B571 ea
EUR 36.00

redox standard solution 500 ml 358 mv

B572 ea
EUR 36.00

Puromycin Solution, 10mg/ml , Cell Cultere Tested

CR026-001 10ml
EUR 201.00

dsGreen Gel Staining Solution, 10000×, 1 mL

20010 1 mL
EUR 134.00

PBS solution pH 7.4, 1000 ml/bottle

10-9402-10 1000 ml/bottle
EUR 161.00

dsGreen Gel Staining Solution, 10000×, 0.5 mL

10010 0.5 mL
EUR 89.00

CometAssay Lysis Solution (2 x 500 mL)

4250-050-01 1 L
EUR 183.00

X-Gal Solution, 40 mg/ml concentrate

X0321-005 5X1ml
EUR 115.00

X-Gal Solution, 40 mg/ml concentrate

X0321-020 20ml
EUR 135.00

Exosome isolation Solution - Cell Media/ Urine (easy version of P100, 95% pure exosome) sample

P100EZ-sample NULL
EUR NULL

Hoechst 33258 *20 mM solution in water*

17525 5 mL
EUR 132.00
  • R-phrase: R20, R21, R68
  • H-Phrase: H303, H313, H340
  • Symbol for dangerous compounds: T
  • UNSPEC Code: 41116134

Hoechst 33342 *20 mM solution in water*

17535 5 mL
EUR 132.00
  • R-phrase: R20, R21, R68
  • H-Phrase: H303, H313, H340
  • Symbol for dangerous compounds: T
  • UNSPEC Code: 41116134

Hoechst 34580 *20 mM solution in water*

17538 100 uL
EUR 219.00
  • R-phrase: R20, R21, R68
  • H-Phrase: H303, H313, H340
  • Symbol for dangerous compounds: T
  • UNSPEC Code: 41116134

Tween® 20, MegaPure? Detergent, 10% Solution

2125-100
EUR 202.00

Tween® 20, MegaPure? Detergent, 10% Solution

2125-50
EUR 147.00

Tween® 20, MegaPure? Detergent, 10% Solution

2125-500
EUR 697.00

Tween® 20, MegaPure? Detergent, 10% Solution

2125-set
EUR 185.00

Pluronic f-127 20% solution in DMSO

59004 1ML
EUR 66.00
Description: Minimum order quantity: 1 unit of 1ML

Sodium dodecyl sulfate, 20% solution in water

GD0990-100ML 100 ml
EUR 54.00

Sodium dodecyl sulfate, 20% solution in water

GD0990-1L 1 l
EUR 126.00

Sodium dodecyl sulfate, 20% solution in water

GD0990-250ML 250 ml
EUR 70.00

Sodium dodecyl sulfate, 20% solution in water

GD0990-2L 2 l
EUR 205.00

FGF-20, human recombinant

4589-20
EUR 277.00

20(R)Ginsenoside?Rh1

N2192-20 20 mg
EUR 514.00
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 224.00
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 514.00
Description: Extracted from Panax ginseng C.A.Mey. roots;Store the product in sealed, cool and dry condition

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

RA806TC-1 10 preps
EUR 554.00
  • Category: MicroRNA Tools

Complete Freund's Adjuvant, 20 mg/ml

7024 20 mg/ml x 5 ml
EUR 393.65
Description: Complete Freund's Adjuvant

Chicken IgY precipitation reagent

85R-1010 4 ml
EUR 554.00
Description: Chicken IgY precipitation reagent for use in immunoassays

PEG Virus Precipitation Kit

K904-200
EUR 1001.00

PEG Virus Precipitation Kit

K904-50
EUR 376.00

PEG Virus Precipitation Kit

K2190-200 200 preparations
EUR 1059.00

PEG Virus Precipitation Kit

K2190-50 50 preparations
EUR 405.00

50ML Protein Precipitation Kit

NAT1254 50ML
EUR 225.00

500 ML HBSS (HANK'S BALANCED SALT SOLUTION), 10X WITH CALCIUM AND MAGNESIUM WITHOUT PHENOL RED AND SODIUM BICARBONATE

20-023-CV 500 mL/pk
EUR 106.00
Description: Media Catalog; Buffered Salt-Salution

100 ML PENICILLIN-STREPTOMYCIN SOLUTION, 50X, 5,000 I.U. PENICILLIN (PER ML) 5,000 µG/ML STREPTOMYCIN

30-001-CI 100 mL/pk
EUR 82.00
Description: Media Catalog; Cell Culture Reagents

Penicillin-Streptomycin 100X Solution, 10, 000 Units/mL, 10, 000 µg/mL

CCM1133-100 100 mL
EUR 64.05
  • Product category: Culture Media

Dispase Solution, 5mg/ml in DMEM/F-12

CA192-020 200 ml
EUR 374.00

G-418. Sulfate, 100mg/ml, Sterile-Filtered Solution

CR008-001 10 ml
EUR 143.00

Acridine orange *10 mg/mL solution in water*

17503 10 mL
EUR 115.00
  • R-phrase: R20, R21, R68
  • H-Phrase: H303, H313, H340
  • Symbol for dangerous compounds: T
  • UNSPEC Code: 41116134

Ethidium bromide [EB, EtBr], 10MG/ml Stock Solution

D0197 5ml
EUR 55.22
  • Product category: Electrophoresis Related/Tracking Dyes

G418 Disulfate Solution (50 mg/ml in Water)

G020-G020-20ML G020-20 mL
EUR 96.00

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.