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August 26, 2020

Western Blotting by Direct Deposition of Reagents onto Capture Membrane

By Danielle

Fast Immunoassay for Microfluidic Western Blotting by Direct Deposition of Reagents onto Capture Membrane

Western blotting is a widely used protein assay platform, but the technique requires long analysis times and multiple manual steps. Microfluidic systems are currently being explored for increased automation and reduction of analysis times, sample volumes, and reagent consumption for western blots. Previous work has demonstrated that proteins separated by microchip electrophoresis can be captured on membranes by dragging the microchip outlet across the membrane.
This process reduces the separation and transfer time of a western blot to a few minutes. To further improve the speed and miniaturization of a complete western blot, a microscale immunoassay with direct deposition of immunoassay reagents has been developed. Flow deposition of antibodies is used to overcome diffusion limited binding kinetics so that the entire immunoassay can be completed in 1 h with detection sensitivity comparable to incubation steps requiring 20 h. The use of low microliter/min flow rates with antibody reagents applied directly and locally to the membrane where the target proteins have been captured, reduced antibody consumption ~30-fold. The complete western blot was applied to the detection of GAPDH and β-Tubulin from A431 cell lysate.

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Resolution Std Toluene in MeOH
RSPEC-EPR002 100ML
EUR 270.26

BioDrop Resolution CFR PC software
SPE6056 EACH
EUR 1744.8

EP Reagent Buffered Acetone Sol.
4000100 1L
EUR 73.2

Hanna Pocket Tester pHECTDSoC Hi Resolution
PHM4132 EACH
EUR 272.4

Hanna Checker pH Tester HI 98103 with 0.1 pH Resolution
PHM4000 EACH
EUR 78

Resolution Std Toluene in MeOH w/ Blank in Cuvettes
RSPEC-EPR001 PK2
EUR 1467.49

RFM340-T digital refractometer 0.000 Bx Resolution Touch Display
REF1148 EACH
EUR 11551.95

REAGENTS HIGH RANGE AMMONIA
HI73325 PK25
EUR 49.2

High Range Ammonia Reagents
HI9373301 EACH
EUR 116.4

RFM330-T digital refractometer 0.0 Bx Resolution Touch Display
REF1144 EACH
EUR 10782.95

Protein A Agarose IP Reagent
abx097210-2ml 2 ml
EUR 326.4

Protein G Agarose IP Reagent
abx097211-2ml 2 ml
EUR 326.4

Protein A&G Agarose IP Reagent
abx097212-2ml 2 ml
EUR 326.4

Agarose, high EEO
GE7747-1 1
EUR 660.6

Agarose, high EEO
GE7747-250 250
EUR 227.8

DNA Loading (6X) Buffer for Agarose and Acrylamide Gels
306-205 5x1 ml
EUR 56.4

Ready-to-Use Ni QR Agarose Beads Buffer Kit
K6563-3 each
EUR 183.6

Cys-link agarose gel coupling buffer kit (5-10 peptides)
110400-CB 1 kit
EUR 270

EZ-DNA Reagents
BS8202 100preps
EUR 105.67

EZ-DNA Reagents
SK8201 100preps
EUR 112.2

Ecl Detection Reagents
RPN2105 EACH
EUR 724.8

Ecl Detection Reagents
RPN3004 EACH
EUR 582

ToxOut™ High Capacity Endotoxin Removal Agarose
7942-5 each Ask for price

Formal Buffered Saline
FBS1 1L
EUR 19.2

Formal Buffered Saline
FBS10 10L
EUR 176.26

Formal Buffered Saline
FBS250 250ML
EUR 24.15

Formal Buffered Saline
FBS5 5L
EUR 54

CryoProtX Mix Reagents 1.5 mL
M-MDSR-61 1.5 ml ml
EUR 37
Description: CryoProtX Mix Reagents 1.5 mL

Alkphos Direct Labelling Reagents
RPN3680 EACH
EUR 813.6

Set of 10 Biolipidure Reagents
Biolipidure-set 10mLx10
EUR 1820.4
Description: Set of 10 Biolipidure Reagents, whose applications include Immunoassays, Western blots, Immunohistochemistry, Turbidimetric assays, Immunochromatography, and Bead based assays. Benefits include: No lot to lot variation, No animal derived materials, Non-specific adsorption suppression, Stabilization of immobilized antibody, Stabilization of enzyme-antibody conjugate, Enzyme-substrate reaction enhancement and aggregation reaction enhancement

Amersham ECL Western Blotting Detection Reagents for 4000cm2 membrane
RPN2106 EACH
EUR 374.4

Amersham ECL Western Blotting Detection Reagents for 1000cm2 membrane
RPN2109 EACH
EUR 188.4

Amersham ECL Western Blotting Detection Reagents for 6000cm2 membrane
RPN2134 1KIT
EUR 572.4

Amersham ECL Western Blotting Detection Reagents for 2000cm2 membrane
RPN2209 EACH
EUR 291.6

CryoProtX Mix Eco Reagents 1.5 mL
M-MDSR-61-ECO 1.5 ml ml
EUR 37
Description: CryoProtX Mix Eco Reagents 1.5 mL

Free Chlorine Reagents 0-5mg/L
WAT1028 EACH
EUR 44.4

10% Neutral Buffered Formalin
NBF105 5L
EUR 142.8

Formalin (10% Neutral Buffered)
FRN125 125 ml
EUR 73.2

Formalin (10% Neutral Buffered)
FRN3800 1 Gal.
EUR 144

Formalin (10% Neutral Buffered)
FRN500 500 ml
EUR 79.2

Formalin (10% Neutral Buffered)
FRN999 1000 ml
EUR 100.8

Agarose for DNA electrophoresis
16-0027-500 500 g
EUR 410.4

buffered in Trehalose
S132-8C1 500 µg
EUR 268.8

BUFFERED PEPTONE WATER
B02-121-10kg 10 kg
EUR 942

BUFFERED PEPTONE WATER
B02-121-2kg 2kg
EUR 250.8

BUFFERED PEPTONE WATER
B02-121-500g 500 g
EUR 111.6

EP Reagent Buffer Sol. pH 3.5
4000600-500ML 500ML
EUR 48

Buffered Pept Water 500
212367 EACH
EUR 136.8

TRIS Buffered Saline (TBS)
GB3555-GB35551l GB3555-1l
EUR 37.7

TRIS Buffered Saline (TBS)
GB3555-GB35552500ml GB3555-2500ml
EUR 75.3

TRIS Buffered Saline (TBS)
GB3555-GB3555500ml GB3555-500ml
EUR 22.7

BUFFERED PEPTONE WATER (ISO)
B02-130-10kg 10 kg
EUR 1148.4

BUFFERED PEPTONE WATER (ISO)
B02-130-2kg 2kg
EUR 296.4

BUFFERED PEPTONE WATER (ISO)
B02-130-500g 500 g
EUR 124.8

Buffered Normal Goat Serum
9066 50 ml
EUR 197.46
Description: Buffered Normal Goat Serum

Buffered Peptone Water (ISO)
MED1030 EACH
EUR 39.6

Buffered Peptone Water (ISO)
MED1032 PK10
EUR 31.2

Buffered Peptone Water (ISO)
MED1374 EACH
EUR 50.4

Formalin, 10% solution, neutral buffered
GE0900-1 1
EUR 22.7

Formalin, 10% solution, neutral buffered
GE0900-2500 2500
EUR 45.2

EP Reagent Acetate Buffer Sol. pH 4.6
4001400 1L
EUR 216

Bradford(Coomassie) Protein Assay Plus Reagents
P7201-050 450ml
EUR 225.6

Phosphate Buffered Saline (PBS)
2113-500 each
EUR 164.4

Phosphate Buffered Saline(PBS)
MED1390 EACH
EUR 33.6

EP Reagent Phosphate Buffer Sol. pH 7.4
4004800 1L
EUR 73.2

EP Reagent Phosphate Buffer Sol. pH 9.0
4008300 1L
EUR 73.2

EP Reagent Phosphate Buffer Sol. pH 8.5
4013300 5L
EUR 272.4

Buffered Glycerol Mounting Media
GMM003 3 ml
EUR 73.2

Buffered Glycerol Mounting Media
GMM030 30 ml
EUR 86.4

Buffered Glycerol Mounting Media
GMM500 500 ml
EUR 219.6

Buffered Glycerol Mounting Media
GMM999 1000 ml
EUR 312

Tris Buffered Saline (TBS) Powder
AR0031 2000mL/pack
EUR 63.6

BUFFERED LISTERIA ENRICHMENT BROTH
B02-131-10kg 10 kg
EUR 950.4

BUFFERED LISTERIA ENRICHMENT BROTH
B02-131-2Kg 2 Kg
EUR 253.2

BUFFERED LISTERIA ENRICHMENT BROTH
B02-131-500g 500 g
EUR 112.8

Biotin Agarose Resin
4BCL-BI-10 10 ml
EUR 493.2

Biotin Agarose Resin
4BCL-BI-5 5 ml
EUR 282

Tris Buffered Saline (10x) pH 7.5
TBD010 10 L
EUR 340.8

Tris Buffered Saline (10x) pH 7.5
TBD500 500 ml
EUR 96

Tris Buffered Saline (10x) pH 7.5
TBD999 1000 ml
EUR 117.6

Tris Buffered Saline (25x) pH 7.4
TBS010 10 L
EUR 340.8

Tris Buffered Saline (25x) pH 7.4
TBS500 500 ml
EUR 96

Tris Buffered Saline (25x) pH 7.4
TBS999 1000 ml
EUR 117.6

Agarose
20-abx186369
  • EUR 610.80
  • EUR 360.00
  • 100 g
  • 25 g

Agarose
20-abx098147
  • EUR 260.40
  • EUR 126.00
  • 100 g
  • 1 g

Agarose
abx082006-100g 100 g
EUR 292.8

Establishment of a candidate equine influenza Florida Clade 2 strain A/eq/Richmond/1/07 horse antiserum as Ph. Eur. Biological Reference Preparation/OIE International Reference Reagent

Equine influenza (EI) is an important respiratory disease of horses, with welfare and economic consequences. Vaccination remains one of the most efficient prevention methods available. Equine influenza virus (EIV) is constantly evolving and consequently EI vaccines need to be updated on a regular basis. In 2010, the World Organisation for Animal Health (OIE) Expert Surveillance Panel (ESP) on EI provided a new recommendation for EI vaccine strain composition, including the incorporation of representative EIV strains of both Florida Clade 1 and Clade 2 sub-lineages (FC1 and FC2, respectively). In this context, the European Pharmacopoeia (Ph. Eur.) – OIE reference panel for EI had to be complemented by an antiserum raised in horses against the FC2 representative EIV strain A/eq/Richmond/1/07.
An international collaborative study was organised and managed by the European Directorate for the Quality of Medicines and HealthCare (EDQM) within the framework of its Biological Standardisation Programme (BSP). The study aimed at evaluating a new candidate reference for use as a common OIE International Standard/Ph. Eur. Biological Reference Preparation (BRP) horse antiserum to FC2 EIV A/equine/Richmond/1/07.
The standard was to be established using the SRH and HI tests for subsequent use in immunogenicity, efficacy and batch potency assay of EI vaccines as a Ph. Eur. BRP (Ph. Eur. monograph 0249) and for use in clinical diagnostic tests as an OIE-approved International Standard Reagent (OIE chapter 3.5.7). The collaborative study confirmed the suitability of the candidate and an SRH titre was assigned. The candidate was adopted as a BRP by the Ph. Eur. Commission and approved by the OIE Biological Standards Commission as an International Standard Serum in November 2017 and February 2018, respectively.

Selective phase transfer reagents (OxP-crowns) for chromogenic detection of nitrates especially ammonium nitrate

Nitrogen and phosphorus-containing ions such as ammonium, nitrates and phosphates are anthropogenic pollutants while ammonium nitrate may be diverted for nefarious purposes in improvised explosive devices. Crown ether-oxoporphyrinogen conjugates (OxP-crowns) are used to selectively detect nitrates, especially their ion pairs with K+ and NH4+, based on ion pair complexation of OxP-crowns under phase transfer conditions.
The presence of phosphate and carbonate lead to deprotonation of OxP-crowns. OxP-1N18C6 is capable of extracting ion pairs with nitrate from aqueous phase leading to a selective chromogenic response. Deprotonation of the OxP moiety leads to [OxP-]-1N18C6[K+] and is promoted by crown ether selective cation binding coupled with hydration of basic oxoanions, which are constrained to remain in the aqueous phase.
This work illustrates the utility of molecular design to exploit partitioning and ion hydration effects establishing selectivity of chromogenic response.

CD45RA Antibody (FITC)
abx139834-100tests 100 tests
EUR 510

CD45RA Antibody (FITC)
abx134000-100Assays 100 Assays
EUR 526.8

CD45RA Antibody (FITC)
abx134000-50Assays 50 Assays
EUR 410.4

CD45RA Antigen (CD45RA/PTPRC) Antibody (FITC)
abx413328-01mg 0.1 mg
EUR 777.6

CD45RA Antigen (CD45RA/PTPRC) Antibody (FITC)
abx415608-01mg 0.1 mg
EUR 610.8

CD45RA Antigen (CD45RA/PTPRC) Antibody (FITC)
abx415609-25ug 25 ug
EUR 326.4

CD45RA Antigen (CD45RA/PTPRC) Antibody (FITC)
abx414287-01mg 0.1 mg
EUR 610.8

CD45RA Antigen (CD45RA/PTPRC) Antibody (FITC)
abx414687-01mg 0.1 mg
EUR 610.8

EZ-DNA Reagents
BS8202 100preps
EUR 105.67

EZ-DNA Reagents
SK8201 100preps
EUR 112.2

Anti-Hu CD45RA FITC
1F-223-T025 25 tests
EUR 146.4

Anti-Hu CD45RA FITC
1F-223-T100 100 tests
EUR 244.8

Ecl Detection Reagents
RPN2105 EACH
EUR 724.8

Ecl Detection Reagents
RPN3004 EACH
EUR 582

REAGENTS HIGH RANGE AMMONIA
HI73325 PK25
EUR 49.2

High Range Ammonia Reagents
HI9373301 EACH
EUR 116.4

CD45RA / CD45RO / CD3 / CD4 Antibody (FITC, PE, PerCP, APC)
abx200640-50tests 50 tests
EUR 828

CD45RA / CD62L / CD3 / CD4 Antibody (FITC, PE, PerCP, APC)
abx200642-50tests 50 tests
EUR 828

CryoProtX Mix Reagents 1.5 mL
M-MDSR-61 1.5 ml ml
EUR 37
Description: CryoProtX Mix Reagents 1.5 mL

Alkphos Direct Labelling Reagents
RPN3680 EACH
EUR 813.6

Set of 10 Biolipidure Reagents
Biolipidure-set 10mLx10
EUR 1820.4
Description: Set of 10 Biolipidure Reagents, whose applications include Immunoassays, Western blots, Immunohistochemistry, Turbidimetric assays, Immunochromatography, and Bead based assays. Benefits include: No lot to lot variation, No animal derived materials, Non-specific adsorption suppression, Stabilization of immobilized antibody, Stabilization of enzyme-antibody conjugate, Enzyme-substrate reaction enhancement and aggregation reaction enhancement

CD3 (FITC) / CD8 (RPE) Combination Antibody
abx410002-50tests 50 tests
EUR 760.8

CD8 (FITC) / CD4 (RPE) Combination Antibody
abx410006-50tests 50 tests
EUR 760.8

CD4 (FITC) / CD8 (RPE) Combination Antibody
abx410011-100tests 100 tests
EUR 744

CD4 (FITC) / CD8 (RPE) Combination Antibody
abx410012-100tests 100 tests
EUR 760.8

CD3 (FITC) / CD8 (RPE) Combination Antibody
abx410015-100tests 100 tests
EUR 744

CD3 (FITC) / CD8 (RPE) Combination Antibody
abx410018-100tests 100 tests
EUR 895.2

CD4 (FITC) / CD8 (RPE) Combination Antibody
abx410019-100tests 100 tests
EUR 895.2

CryoProtX Mix Eco Reagents 1.5 mL
M-MDSR-61-ECO 1.5 ml ml
EUR 37
Description: CryoProtX Mix Eco Reagents 1.5 mL

Free Chlorine Reagents 0-5mg/L
WAT1028 EACH
EUR 44.4

CD4 (FITC) / CD8 (RPE) / CD3 (RPE-Cy5) Combination Antibody
abx410020-50tests 50 tests
EUR 744

CD45RA FITC Monoclonal Antibody (Clone HI100)
6959-100 each
EUR 379.2

CD45RA FITC Monoclonal Antibody (Clone HI100)
6959-25 each
EUR 196.8

Bradford(Coomassie) Protein Assay Plus Reagents
P7201-050 450ml
EUR 225.6

CD8/CD38 (CD8/CD38) Antibody (FITC, PE)
abx200605-50tests 50 tests
EUR 510

CD8 / CD56 Antibody (FITC, PE)
abx200606-50tests 50 tests
EUR 510

CD3 / CD8 Antibody (FITC, PE)
abx200597-50tests 50 tests
EUR 510

CD4 / CD8 Antibody (FITC, PE)
abx200600-50tests 50 tests
EUR 510

CD8 / CD4 Antibody (FITC, PE)
abx200604-50tests 50 tests
EUR 510

CD4 / CD8 Antibody (FITC / PE)
abx159408-100ul 100 ul
EUR 427.2

CD45RA Antigen (CD45RA/PTPRC) Antibody (RPE)
abx413330-100tests 100 tests
EUR 861.6

CD45RA Antigen (CD45RA/PTPRC) Antibody (RPE)
abx415610-100tests 100 tests
EUR 710.4

CD45RA Antigen (CD45RA/PTPRC) Antibody (RPE)
abx414690-100tests 100 tests
EUR 710.4

Amersham ECL Western Blotting Detection Reagents for 4000cm2 membrane
RPN2106 EACH
EUR 374.4

Amersham ECL Western Blotting Detection Reagents for 1000cm2 membrane
RPN2109 EACH
EUR 188.4

Amersham ECL Western Blotting Detection Reagents for 6000cm2 membrane
RPN2134 1KIT
EUR 572.4

Amersham ECL Western Blotting Detection Reagents for 2000cm2 membrane
RPN2209 EACH
EUR 291.6

CD4 / CD8 / CD3 Antibody (FITC, PE, PE-Cy5)
abx200628-50tests 50 tests
EUR 710.4

CD4 / CD8 / CD3 Antibody (FITC / PE / PE-Cy5)
abx159409-100ul 100 ul
EUR 427.2

CD3 / CD8 / CD45 Antibody (FITC, PE, PerCP)
abx201076-50tests 50 tests
EUR 678

HLA-DR / CD8 Antibody (FITC, PE)
abx200621-50tests 50 tests
EUR 510

CD4 / CD8 / CD3 Antibody (FITC, PE, PerCP)
abx200629-50tests 50 tests
EUR 678

CD8 FITC Antibody
20-abx137160
  • EUR 560.40
  • EUR 777.60
  • 0.5 mg
  • 1 mg

CD8 antibody (FITC)
61-001103F 50 ug
EUR 289.2
Description: Mouse monoclonal CD8 antibody (FITC)

CD8 antibody (FITC)
61R-1046 100 ug
EUR 198
Description: Rat monoclonal CD8 antibody (FITC)

CD8 antibody (FITC)
61R-1047 100 ug
EUR 230.4
Description: Rat monoclonal CD8 antibody (FITC)

CD8 antibody (FITC)
61R-1872 500 ug
EUR 664.8
Description: Mouse monoclonal CD8 antibody (FITC)

CD8 antibody (FITC)
61R-CD8bHUFT 100 tests
EUR 327.6
Description: Mouse monoclonal CD8 antibody (FITC)

CD8 antibody (FITC)
61R-CD8dFEFT 500 ug
EUR 678
Description: Mouse monoclonal CD8 antibody (FITC)

CD45RA/CD45RO/CD3/CD8
45RAF245ROPE3PP18A3-50T 50 test
EUR 730.8

CD45RA/CD62L/CD3/CD8
45RAF262LPE3PP18A3-50T 50 test
EUR 730.8

CD4 (FITC) / CD25 (RPE) Combination Antibody
abx410008-50tests 50 tests
EUR 760.8

CD4 (FITC) / CD25 (RPE) Combination Antibody
abx410013-100tests 100 tests
EUR 744

Mouse Monoclonal Anti-Rat CD45RA/B FITC (Clone OX-33) (mouse IgG1)
RCD045RAB-F 100 tests
EUR 562.8

CD3 / CD8 / CD45 / CD4 Antibody (FITC, PE, PE-Cy5, APC)
abx200638-50tests 50 tests
EUR 878.4

CD79A (FITC) / CD5 (RPE) Combination Antibody
abx410009-50tests 50 tests
EUR 811.2

T7 gRNA SmartNuclease Synthesis Kit (includes CAS510A-1 & T7 IVT synthesis reagents)
CAS510A-KIT 1 Kit
EUR 724

CD3 (FITC) / CD4 (RPE) Combination Antibody
abx410001-50tests 50 tests
EUR 760.8

CD3 (FITC) / CD4 (RPE) Combination Antibody
abx410003-50tests 50 tests
EUR 760.8

CD3 (FITC) / CD4 (RPE) Combination Antibody
abx410004-50tests 50 tests
EUR 760.8

CD3 (FITC) / CD4 (RPE) Combination Antibody
abx410014-100tests 100 tests
EUR 744

CD3 (FITC) / CD4 (RPE) Combination Antibody
abx410017-100tests 100 tests
EUR 895.2

Anti-Hu CD8 FITC
1F-207-T025 25 tests
EUR 146.4

Anti-Hu CD8 FITC
1F-207-T100 100 tests
EUR 244.8

Mouse Monoclonal Anti-Human CD4 FITC/CD8 PE
CD04-D3-50 50 tests
EUR 781.2

Mouse Monoclonal Anti-Human CD8 FITC/CD2 PE
CD08-D1-50 50 tests
EUR 781.2

Mouse Monoclonal Anti-Human CD8 FITC/CD4 PE
CD08-D2-50 50 tests
EUR 781.2

Mouse Monoclonal Anti-Human CD3 FITC/CD8 PE
CD03-D5-50 50 tests
EUR 781.2

Mouse Monoclonal Anti-Human CD3 FITC/CD8 PE/CD45 PE-Cy5
CD03-T3-100 50 Tests
EUR 708

Mouse Monoclonal Anti-Human CD4 FITC/CD8 PE/CD45 PE-Cy5
CD04-T2-100 50 Tests
EUR 708

Mouse Monoclonal Anti-Human CD45 FITC/CD8 PE/CD3 PE-Cy5
CD45-T4-100 50 Tests
EUR 708

Exo-Flow 2.0 Basic Kit without antibody (Streptavidin beads + reagents) - for Serum or Plasma
EXOFLOW2-BASICA-SP 30 rxn
EUR 1201.2

Mouse Monoclonal Anti-Human CD4 FITC/CD8 PE/CD3 PE-Cy5
CD04-T1-100 50 Tests
EUR 708

Mouse Monoclonal Anti-Human CD8 FITC/CD4 PE/CD3 PE-Cy5
CD08-T1-100 50 Tests
EUR 708

cAMP Biotrak(TM) EIA; (Non-Acetylation Protocol; Lysis Reagents not Included); Cytiva; RPN2251
GERPN2251 EACH
EUR 685.2

CD8 beta Antibody (FITC)
20-abx200907
  • EUR 343.20
  • EUR 292.80
  • EUR 477.60
  • 100 ug
  • 25 ug
  • 500 ug

Annexin V-FITC Reagent
1001-1000 each
EUR 1495.2

Annexin V-FITC Reagent
1001-200 each
EUR 529.2

CD8 alpha Antibody (FITC)
20-abx200899
  • EUR 343.20
  • EUR 292.80
  • 100 ug
  • 25 ug

MPO (FITC) / CD3 (RPE) Combination Antibody
abx410010-50tests 50 tests
EUR 543.6

Exo-Flow 2.0 Basic Kit without antibody (Streptavidin beads + reagents) - for Tissue Culture Media
EXOFLOW2-BASICA-TC 30 rxn
EUR 1201.2

CD3 (FITC) / HLA2 (RPE) Combination Antibody
abx410007-50tests 50 tests
EUR 760.8

Mouse anti-Human CD3/CD8, FITC/PE Conjugated mAb
28235-20Tests 20 Tests
EUR 302.4

Mouse anti-Human CD4/CD8, FITC/PE Conjugated mAb
28236-20Tests 20 Tests
EUR 302.4

CD45RA Antibody (PE)
abx200320-100tests 100 tests
EUR 727.2
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