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Table 2 Details of primary and secondary antibodies used during western blotting

From: Putative molecular mechanism underlying sperm chromatin remodelling is regulated by reproductive hormones

Antibodies MW (Kd) Antibody dilutions
Primary Secondary Ref (primaries)
UBE1 (N-20) 110 1:200 (Goat) 1:50,000 (Sigma) Santa Cruz 47555
E217Kb/UBE2D2 17 1:1,000 (Rb) 1:50,000 (Sigma) S.S.Wing (UBC4) (gift)
LASU1 /URE-B1/E3 485 1:1,000 (Rb) 1:50,000 (Sigma) S.S.Wing (HUWE1) (gift)
CDYL (F-15) 66 1:50 (Goat) 1:20,000 (Sigma) Santa Cruz 34147
BRDT/brd6 75 1:100 (Goat) 1:5,000 (s.c) Everest EB05796
HDAC1(C-19) 52 1:100 (Goat) 1:50,000 (Sigma) Santa Cruz 6298
TOPO IIβ (H-286) 180 1:200 (Rb) 1:5,000 (s.c) Santa Cruz 13059
PEM (M-15) 21-24 1:50 (Goat) 1:4,000 (s.c) Santa Cruz 21650
Proteasomes purified fish 24-31 1:3,000 (Ms Mab) 1:50,000 (Sigma) T. Tokumoto (GC3α) (gift)
20S Proteasomeα1(C7) 24-31 1:200 (Ms Mab) 1:5,000 (s.c) Santa Cruz 166073
20S Proteasomeα5(H-53) 24-31 1:250 (Rb) 1:5,000 (s.c) Santa Cruz 67342
20S Proteasomeα7(MCP72) 24-31 1:500 (Ms MAb) 1:50,000 (Sigma) Santa Cruz 58417
H1 17 1:20 (Ms Mab) 1:4,000 (s.c) Abcam 62884
H2A somatic 14 1:20 (Rb) 1:20,000 (Sigma) Biovision 3621-100
H2Bsomatic 14 1:3,000 (Rb) 1:4,000 (s.c) Epitomics 1847-1
H3 somatic 15 1:1,000 (Rb) 1:50,000 (Sigma) Santa Cruz 8654R
tH2B 15 1:250 (Rb) 1:25,000 (Sigma) Millipore 07680
H4 11 1:20 (Rb) 1:20,000 (Sigma) Biovision 3624-100
H2AUb 25 1:65 (Ms Mab/IgM) 1:8,500 (anti-IgM:Sigma) Millipore 05678
H2BUb 25 1:100 (Mab) 1:4,000 (s.c) Millipore 051312
Ub histones 12-225 1:50 (Rb) 1:4,000 (s.c.) Abcam/UbB/7780
Ub histones FK1 12-225 1:100 (MsMab/IgM) 1:9,000 (anti-IgM:Sigma) Enzo/FK1 PW8805
H2AK5 14 1:65 (Rb) 1:4,000 (s.c) Cell Signalling 2576
H2BK5 14 1:200 (Rb) 1:4,000 (s.c) Cell Signalling 2574
H4K5 11 1:5,000 (Rb MAb) 1:2,000 (s.c) Epitomics 1808-1
H4K12 11 1:200 (Rb) 1:20,000 (Sigma) Abcam 1761
   1:200 (Rb) 1:4,000 (s.c) Cell Signalling 2591
H4K Penta 10 1:30 (Rb) 1:20,000 (Sigma) Millipore 06946
  1. ~MW is denoted as Kd. GC3α MAb is used to detect Proteasomes. Ubiquitin antibody and FK1 MAb IgM antibody are used to detect ubiquitylated and polyubiquitylated histones respectively. H4 Penta is used to detect penta-acetylated H4.