Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|
|---|---|---|---|---|---|
| Chemical compound, drug |
Trizma base | Sigma aldrich | Cat# T1503-5KG | ||
| Chemical compound, drug |
Boric acid | Sigma Aldrich | Cat# B6768-5KG | ||
| Chemical compound, drug |
EDTA | Sigma Aldrich | Cat# EDS-500G | ||
| Chemical compound, drug |
DTT | Thermo Fisher Scientific |
Cat# BP17225 | ||
| Chemical compound, drug |
HEPES | Sigma Aldrich | Cat# H4034-100G | ||
| Chemical compound, drug |
Proteinase K | Thermo Fisher Scientific |
Cat# FEREO0491 | ||
| Chemical compound, drug |
Magnesium chloride hexahydrate |
Sigma Aldrich | Cat# M2670-500G | ||
| Chemical compound, drug | Potassium chloride | Sigma Aldrich | Cat# P9541-500G | ||
| Chemical compound, drug |
Sodium chloride | Thermo Fisher Scientific |
Cat# S271-3 | ||
| Chemical compound, drug | Sodium acetate | Sigma Aldrich | Cat# S2889-250G | ||
| Chemical compound, drug |
Tryptone | Thermo Fisher Scientific | Cat# BP1421-500 | ||
| Chemical compound, drug | SDS micropellets | Thermo Fisher Scientific |
Cat# BP8200-500 | ||
| Chemical compound, drug | Agarose | Thermo Fisher Scientific |
Cat# BP1356-500 | ||
| Chemical compound, drug | PMSF | Calbiochem | Cat# 7110–5 GM | ||
| Chemical compound, drug | Triton X-100 | Sigma Aldrich | Cat# T8787-250ML | ||
| Chemical compound, drug | Lithium chloride | Sigma Aldrich | Cat# L4408-500G | ||
| Chemical compound, drug | Lithium acetate dihydrate |
Sigma Aldrich | Cat# CAS6108-17-4 | ||
| Chemical compound, drug |
Tween 20 | Thermo Fisher Scientific | Cat# CAS9005-64-5 | ||
| Chemical compound, drug |
Peptone | RPI Research Products |
Cat# P20240-1000.0 | ||
| Chemical compound, drug | Leupeptin | RPI Research Products |
Cat# L22035-0.025 | ||
| Chemical compound, drug |
Aprotinin | RPI Research Products |
Cat# A20550-0.05 | ||
| Chemical compound, drug |
Pepstatin | RPI Research Products | Cat# P30100-0.025 | ||
| Chemical compound, drug |
N-Oxalylglycine | Sigma Aldrich | Cat# O9390-10MG | ||
| Chemical compound, drug |
α-Ketoglutaric acid disodium salt dihydrate |
Sigma Aldrich | Cat# 75892–25G | ||
| Chemical compound, drug | SuperSignal West Pico PLUS Chemiluminescent Substrate |
Thermo Scientific | Cat# B2162617 | ||
| Chemical compound, drug | Trichloroacetic acid | Sigma Aldrich | Cat# T0699-100ML | ||
| Chemical compound, drug |
Phenol:chloroform: isoamyl alcohol |
Sigma Aldrich | Cat# P3803-100ML | ||
| Chemical compound, drug | L-ascorbic acid | Fisher Chemical | Cat# C6H8O6 | ||
| Chemical compound, drug |
Coblat(II) chloride hexahydrate |
Sigma Aldrich | Cat# 255599–100G | ||
| Chemical compound, drug |
Ammonium iron (II) sulfate hexahydrate |
ACROS ORGANICS | Cat# 423721000 | ||
| Chemical compound, drug | Formaldehyde | Sigma Aldrich | Cat# 252549–500 ML | ||
| Chemical compound, drug |
Acetonitrile | OmniSolv | Cat# AX0156-1 | ||
| Chemical compound, drug |
Trifluoroacetic acid | Fisher Chemical | Cat# A116−10 × 1 AMP | ||
| Chemical compound, drug | Dimethyl pimelimidate | Thermo Scientific | Cat# 21667 | ||
| Chemical compound, drug | Ammonium hydroxide solution | Sigma Aldrich | Cat# 221-228-100ML-A | ||
| Chemical compound, drug | Ethanolamine | Sigma Aldrich | Cat# 411000–100 ML | ||
| Chemical compound, drug |
Ethyleneglycol bis succinimidylsuccinate | Thermo Scientific | Cat# 21565 | ||
| Chemical compound, drug | Glycogen | Sigma Aldrich | Cat# 10901393001 | ||
| Chemical compound, drug | GelGreen Nucleic Acid Stain | BioTium | Cat# 41004 | ||
| Chemical compound, drug |
30% Acrylamide/ Bis solution, 37.5:1 |
Bio-Rad | Cat# 1610158 | ||
| Chemical compound, drug | Ammonium persulfate | Bio-Rad | Cat# A3678-100G | ||
| Chemical compound, drug | TEMED | Sigma Aldrich | Cat# T9281-50ml | ||
| Chemical compound, drug | Hydrochloric acid | Thermo Fisher Scientific | Cat# A14-500 | ||
| Chemical compound, drug | Agar | Sigma Aldrich | Cat# A1296-1KG | ||
| Chemical compound, drug | IPTG | Thermo Fisher Scientific | Cat# BP1755-10 | ||
| Antibody | Anti-H3K9me2 (Mouse monoclonal) | Abcam | Cat# ab1220 RRID:AB_449854 |
IF (1:1000), WB (1:2500) | |
| Antibody | Anti-H3K9me3 (Rabbit polyclonal) |
Abcam | Cat# ab8898 RRID:AB_306848 |
IF (1:1000), WB (1:5000) | |
| Antibody | Anti-H3 (Rabbit polyclonal) | Abcam | Cat# ab1791 RRID:AB_302613 |
IF (1:1000), WB (1:5000) | |
| Antibody | Anti-H3K4me3 (Rabbit polyclonal) |
Abcam | Cat# ab8580 RRID:AB_2827504 |
IF (1:1000), WB (1:1000) | |
| Antibody | Monoclonal ANTI- FLAG M2 Antibody |
Abcam | Cat# F1804-5MG RRID:AB_262044 |
IF (1:5000), WB (1:5000) | |
| Antibody | THE V5 Tag Antibody | GenScript | Cat# A01724-100 RRID:AB_2827501 |
IF (1:1000), WB (1:5000) | |
| Antibody | Anti-MBP (Mouse monoclonal) |
New England Biolabs | Cat# E8032S RRID:AB_2827502 |
WB (1:5000) | |
| Antibody | Anti-Swi6 (Rabbit polyclonal) |
Custom | WB (1:2500) | ||
| Peptide, recombinant protein | H3K9me3 (1603 Da) | New England Peptide | Custom | H2N-ARTKQTAR(K9me3) STGGKA-amide |
|
| Peptide, recombinant protein | H3K9me0 (1560 Da) | New England Peptide | Custom | ARTKQTKARKSTGGKA-amide | |
| Peptide, recombinant protein |
H3K9(me2) (2751 Da) | Anaspec peptide | Cat# AS-64359 | H-ARTKQTARK(ME2)STGGKAPPKQLAGGK(biotin)-OH | |
| Peptide, recombinant protein |
H3K9(me3) (2766 Da) | Anaspec peptide | Cat# AS-64360 | H-ARTKQTARK(ME3)STGGKAPPKQLAGGK(biotin)-OH | |
| Peptide, recombinant protein | H3 (2722 Da) | Anaspec peptide | Cat# AS-61702 | H-ARTKQTARKSTGGKAP
PKQLAGGK(biotin)-OH |
|
| Other | Dynabeads Protein G | Thermo Fisher Scientific |
LOT# 00448217 | ||
| Other | Dynabeads M-280 Streptavidin | Thermo Fisher Scientific | LOT# 00448388 | ||
| Other | Dynabeads Protein A | Thermo Fisher Scientific |
LOT# 00689576 | ||
| Other | ANTI-FLAG M2 Affinity Gel |
Sigma Aldrich | LOT# A2220-5ML | ||
| Other | Amylose Resin | New England Biolabs |
LOT# E8021L | ||
| Other | Pierce Gutathione Agarose, 100 ml | Thermo Scientific | LOT# R1241698 | ||