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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.
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Clinical Epigenetics

ISSN: 1868-7083

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