Blocking buffer (ELISA) |
|
|
10 % FBS in PBS |
Cell culture flask T25 |
Greiner Bio-One |
690175 |
we use standard CELLSTAR filter cap cell culture flasks; alternatively use suspension culture flask (690195 ) |
Cell culture flask T75 |
Greiner Bio-One |
658175 |
we use standard CELLSTAR filter cap cell culture flasks; alternatively use suspension culture flask (658195) |
Cell culture flask T175 |
Greiner Bio-One |
661175 |
we use standard CELLSTAR filter cap cell culture flasks; alternatively use suspension culture flask (661195) |
Centrifugal concentrator MWCO 10 kDa |
Sartorius |
VS2001 |
Vivaspin 20 centrifugal concentrator |
Centrifugal protein concentrator MWCO 100 kDa, 5 – 20 ml |
Thermo Fisher Scientific |
88532 |
Pierce Protein Concentrator, PES 5 -20 ml; we use the Pierce Concentrator 150K MWCO 20mL (catalog number 89921), which is however no longer available |
Centrifuge bottles |
Nalgene |
525-2314 |
PPCO (polypropylene copolymer) with PP (polypropylene) screw closure, 500 ml; obtained from VWR, Germany |
Coating buffer (ELISA) |
|
|
0.1 M sodium bicarbonate (NaHCO3) in H2O (pH 9.6) |
Desalting columns MWCO 7 kDa |
Thermo Fisher Scientific |
89891 |
Thermo Scientific Zeba Spin Desalting Columns, 7K MWCO, 5 mL |
Detection reagent ELISA (HRPO substrate) |
Sigma-Aldrich/Merck |
T8665-100ML |
3,3′,5,5′-Tetramethylbenzidine (TMB) liquid substrate system |
ELISA 96-well plate |
Thermo Fisher Scientific |
442404 |
MaxiSorp Nunc-Immuno Plate |
Fetal calf serum |
PAN-BIOtech |
P40-47500 |
FBS Good forte |
ISF-1 medium |
Biochrom/bioswisstec |
F 9061-01 |
|
NLDC-145 hybridoma |
ATCC |
HB-290 |
if not already at hand, the hybridoma cells can be acquired from ATCC |
Ovalbumin |
Hyglos (via BioVendor) |
321000 |
EndoGrade OVA ultrapure with <0.1 EU/mg |
Penicillin/Streptomycin |
Thermo Fisher Scientific |
15140122 |
Gibco Penicillin/Streptomycin 10.000 U/ml; alternatively Gibco Penicillin/Streptomycin 5.000 U/ml (15070-063) can be used |
PETG polyethylene terephthalate glycol cell culture roller bottles |
Nunc In Vitro |
734-2394 |
standard PDL-coated, vented (1.2X), 1050 cm², 100 – 500 ml volume; obtained from VWR, Germany |
pH indicator strips |
Merck |
109535 |
pH indicator strips 0-14 |
Polyclonal goat αrat-IgG+IgM-HRPO antibody (ELISA) |
Jackson ImmunoResearch |
112-035-068 |
obtained from Dianova, Germany; used at 1:2000 for ELISA |
Polyclonal rabbit αOVA (ELISA) |
Abcam |
ab181688 |
used at 3 ng/µl |
Protein G Sepharose column |
Merck/Millipore |
P3296 |
5 ml Protein G Sepharose, Fast Flow are packed onto an empty column PD-10 (Merck, GE 17-0435-01) |
Rubber plug |
Omnilab |
5230217 |
DEUTSCH & NEUMANN rubber stoppers (lower Φ 17 mm; upper Φ 22 mm) |
Silicone tube |
Omnilab |
5430925 |
DEUTSCH & NEUMANN (inside Φ 1 mm; outer Φ 3 mm) |
Slim-Fast |
|
|
we have used regular Slim-Fast Chocolate freely available at the pharmacy as in this western blot approach it yielded better results than milk powder |
Stopping solution (ELISA) |
|
|
1M H2SO4 |
Sulfo-SMCC |
Thermo Fisher Scientific |
22322 |
Pierce Sulfo-SMCC Cross-Linker; alternatively use catalog number A39268 (10 x 2 mg) |
Syringe filter unit 0.22 µm |
Merck/Millipore |
SLGV033RS |
Millex-GV Syringe Filter Unit, 0.22 µm, PVDF, 33 mm, gamma sterilized |
Syringe 10 ml |
Omnilab |
|
Disposable syringes Injekt® Solo B.Braun |
Sterican® cannulas |
B. Braun |
|
Sterican® G 20 x 1 1/2""; 0.90 x 40 mm; yellow |
TCEP-HCl |
Thermo Fisher Scientific |
A35349 |
|
Tubing connector |
Omnilab |
|
Kleinfeld miniature tubing connectors for silicone tube |