Histone H3 Nuclear Loading Control Polyclonal Antibody is a Rabbit antibody targeting Histone H3 Nuclear Loading Control. Histone H3 Nuclear Loading Control Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Fruit Fly,Rabbit,Cow,Pig)

1. Paraformaldehyde-fixed, paraffin embedded (mouse testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
2. Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (mouse liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (mouse liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
5. Blank control: Mouse spleen cells (blue). Primary Antibody: Rabbit Anti-Histone H3/HIST3H3 antibody (TMAB-00866), Dilution: 1 μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit Igg (orange),used under the same conditions); Secondary Antibody: Goat anti-rabbit IgG-Pe (white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min). Primary antibody (TMAB-00866, 1 μg/1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 10% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice.
6. Paraformaldehyde-fixed, paraffin embedded (human colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded (rat pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded (mouse pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
9. Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
10. Paraformaldehyde-fixed, paraffin embedded (mouse kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
11. Paraformaldehyde-fixed, paraffin embedded (human laryngeal carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3HIST3H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
12. Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3HIST3H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
13. Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3HIST3H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
14. Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
15. Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
16. Paraformaldehyde-fixed, paraffin embedded (Human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
17. Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
18. Paraformaldehyde-fixed, paraffin embedded (mouse kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
19. Paraformaldehyde-fixed, paraffin embedded (rat colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
20. Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
21. Paraformaldehyde-fixed, paraffin embedded (mouse testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
22. Paraformaldehyde-fixed, paraffin embedded (rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
23. Paraformaldehyde-fixed, paraffin embedded (human liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
24. Paraformaldehyde-fixed, paraffin embedded (human brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nucleus staining.
25. Blank control: K562. Primary Antibody (green line): Rabbit Anti-Histone H3/HIST3H3 antibody (TMAB-00866)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-PE
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
26. 25 μg total protein per Lane of various lysates probed with Histone H3 polyclonal antibody, unconjugated (TMAB-00866) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at RT for 60 min.
27. Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:1000 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
28. Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (TMAB-00866) at 1:1000 overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

WB: 1:5000-50000; IHC-P: 1:500-2000; IHC-Fr: 1:500-2000; IF: 1:500-2000; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human Histone H3

Human

ChIP antibodies,Methylated