Production of Elastin-like Protein Hydrogels for Encapsulation - JoVE

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Journal of Visualized Experiments

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Materials List for:

Production of Elastin-like Protein Hydrogels for Encapsulation and Immunostaining of Cells in 3D 1

2

1

2

Bauer L. LeSavage* , Nicholas A. Suhar* , Christopher M. Madl , Sarah C. Heilshorn 1

Department of Bioengineering, Stanford University

2

Department of Materials Science and Engineering, Stanford University

*

These authors contributed equally

Correspondence to: Sarah C. Heilshorn at [email protected] URL: https://www.jove.com/video/57739 DOI: doi:10.3791/57739

Materials Name

Company

Catalog Number

10 cm Petri Dishes

Thermo Fisher Scientific

FB0875713

70% Ethanol

RICCA Chemical

2546.70-1

Ammonium Sulfate

Sigma-Aldrich

A3920-500G

Ampicillin

Thermo Fisher Scientific

BP1760-25G

Bacto Agar

Thermo Fisher Scientific

9002-18-0

Comments

Elastin-Like Protein Expression and Purification

BL21(DE3)pLysS Competent Cells Invitrogen

C606003

Chloramphenicol

Amresco

0230-100G

Deoxyribonuclease I from bovine pancreas

Sigma-Aldrich

DN25

EDTA disodium salt, dihydrate

Thermo Fisher Scientific

O2793-500

Glycerol

Thermo Fisher Scientific

BP229-4

Isopropanol

Thermo Fisher Scientific

A451-4

Isopropyl β-D-1thiogalactopyranoside (IPTG)

Thermo Fisher Scientific

BP1755-10G

Luria Broth

EMD Millipore

1.10285.5007

Parafilm

VWR

52858-000

Phenylmethanesulfonyl fluoride (PMSF)

MP Biomedicals

195381

Sodium Chloride

Thermo Fisher Scientific

BP358-212

Sodium Hydroxide

Sigma-Aldrich

S 8045-1KG

Syringe Filter Unit (0.22 μm)

Millipore

SLGP033RB

Terrific Broth

Millipore

71754-4

Tris Base

Thermo Fisher Scientific

BP152-1

0.22 μm syringe filters

Millipore

SLGV004SL

0.5 mm thick silicone sheet

Electron Microscopy Science

70338-05

24-well tissue culture plates

Corning

353047

Disposable Biopsy Punch (2 mm)

Integra Miltex

33-31

Disposable Biopsy Punch (4 mm)

Integra Miltex

33-34

Disposable Biopsy Punch (5 mm)

Integra Miltex

33-35

Cell Encapsulation in 3D ELP Hydrogels

Copyright © 2018 Journal of Visualized Experiments

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Journal of Visualized Experiments

www.jove.com

Dulbecco’s phosphate buffered saline (DPBS)

Corning

21-031-CM

No. 1 12 mm glass coverslips

Thermo Fisher Scientific

12-545-80

Tetrakis(hydroxymethyl)phosphoniumSigma-Aldrich chloride (THPC)

404861-100ML

0.5% Tryspin/EDTA

Thermo Fisher

15400054

16% (w/v) Paraformaldehyde (PFA)

Electron Microscopy Sciences

15701

Bovine Serum Albumin (BSA)

Roche

3116956001

DAPI (4',6-Diamidino-2Phenylindole, Dihydrochloride)

Molecular Probes

D1306

Donkey Serum

Lampire Biological Labs

7332100

Goat anti-mouse Secondary Antibody (AF488)

Molecular Probes

A-11017

Goat anti-rabbit Secondary Antibody (AF546)

Molecular Probes

A-11071

Goat Serum

Gibco

16210-072

Mouse Nestin Primary Antibody

BD Pharmingen

556309

Mouse Sox2 Primary Antibody

Cell Signaling Technology

23064S

Nail Polish

Electron Microscopy Sciences

72180

Triton X-100

Sigma-Aldrich

X100-100ML

Vectashield Hardset Mounting Medium

Vector Labs

H-1400

Immunocytochemistry of Cells in 3D ELP Hydrogels

Copyright © 2018 Journal of Visualized Experiments

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Production of Elastin-like Protein Hydrogels for Encapsulation - JoVE

Journal of Visualized Experiments www.jove.com Materials List for: Production of Elastin-like Protein Hydrogels for Encapsulation and Immunostainin...

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