Trichostatin A treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 83041-1-RR (Acetyl-Histone H2A (Lys9) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with Beta Actin Monoclonal antibody (66009-1-Ig) as loading control.
Trichostatin A treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 83041-1-RR (Acetyl-Histone H2A (Lys9) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with Beta Actin Monoclonal antibody (66009-1-Ig) as loading control.
IHC staining of mouse lung using 83041-1-RR
Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 83041-1-RR (Acetyl-Histone H2A (Lys9) antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 83041-1-RR (Acetyl-Histone H2A (Lys9) antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IF Staining of HeLa using 83041-1-RR
Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Acetyl-Histone H2A (Lys9) antibody (83041-1-RR, Clone: 230349B1 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).
Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Acetyl-Histone H2A (Lys9) antibody (83041-1-RR, Clone: 230349B1 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).
IF Staining of HeLa using 83041-1-RR
Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Acetyl-Histone H2A (Lys9) antibody (83041-1-RR, Clone: 230349B1 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).
Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Acetyl-Histone H2A (Lys9) antibody (83041-1-RR, Clone: 230349B1 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).
IF Staining of HeLa using 83041-1-RR
Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Acetyl-Histone H2A (Lys9) antibody (83041-1-RR, Clone: 230349B1) at dilution of 1:150 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).
Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Acetyl-Histone H2A (Lys9) antibody (83041-1-RR, Clone: 230349B1) at dilution of 1:150 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).
Dot Blot experiment of peptide using 83041-1-RR
Dot blot analysis was used to confirm the specificity of Acetyl-Histone H2A (Lys9) antibody. Acetylated peptides were spotted onto NC and probed with antibody at 1 µg/ml.The amount of peptide (μg/mL) spotted is indicated next to each row.Column 1: H2AEK9Ac. Column 2: Unmodified H2AEK9. Column 3: H2AEK5Ac. Column 4: Unmodified H2AEK5. Column 5: H2AEK13Ac. Column 6: Unmodified H2AEK13. Column 7: H2AEK15Ac. Column 8: Unmodified H2AEK15. Column 9: Blank(PBS).
Dot blot analysis was used to confirm the specificity of Acetyl-Histone H2A (Lys9) antibody. Acetylated peptides were spotted onto NC and probed with antibody at 1 µg/ml.The amount of peptide (μg/mL) spotted is indicated next to each row.Column 1: H2AEK9Ac. Column 2: Unmodified H2AEK9. Column 3: H2AEK5Ac. Column 4: Unmodified H2AEK5. Column 5: H2AEK13Ac. Column 6: Unmodified H2AEK13. Column 7: H2AEK15Ac. Column 8: Unmodified H2AEK15. Column 9: Blank(PBS).
ChIP experiment of HeLa using 83041-1-RR
Chromatin was prepared from HeLa cells, cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 15 µg of cross-linked chromatin, 5 µg of Acetyl-Histone H2A (Lys9) (83041-1-RR) or 5 ug of Normal Rabbit IgG (98136-1-RR), and 20 µl of Protein A Magarose Beads. The immunoprecipitated DNA was quantified by real-time PCR.
Chromatin was prepared from HeLa cells, cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 15 µg of cross-linked chromatin, 5 µg of Acetyl-Histone H2A (Lys9) (83041-1-RR) or 5 ug of Normal Rabbit IgG (98136-1-RR), and 20 µl of Protein A Magarose Beads. The immunoprecipitated DNA was quantified by real-time PCR.
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mouse lung tissue Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
Positive IF/ICC detected in
HeLa cells
Positive Dot Blot detected in
peptide
Positive ChIP detected in
HeLa cells
Recommended dilution
Application
Dilution
Western Blot (WB)
WB : 1:2000-1:10000
Immunohistochemistry (IHC)
IHC : 1:250-1:1000
Immunofluorescence (IF)/ICC
IF/ICC : 1:200-1:800
DOT BLOT
DOT BLOT : 1:10-1:100
Chromatin immunoprecipitation (ChIP)
CHIP : 1:10-1:100
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.
Product Information
83041-1-RR targets Acetyl-Histone H2A (Lys9) in WB, IHC, IF/ICC, ChIP, Dot Blot, ELISA applications and shows reactivity with human, mouse samples.
PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
Storage Conditions
Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. 20ul sizes contain 0.1% BSA.
Background Information
Histone H2A is a core component of nucleosome. Histone variants contribute to chromatin complexity by creating specialized nucleosomes. Within nucleosomes, either one canonical H2A or both of them can be exchanged with a particular variant (heterotypic and homotypic nucleosomes, respectively), and such changes can have profound influences on nucleosome stability and biological outcome.
Protocols
Product Specific Protocols
WB protocol for Acetyl-Histone H2A (Lys9) antibody 83041-1-RR
Chromatin was prepared from HeLa cells, cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 15 µg of cross-linked chromatin, 5 µg of Acetyl-Histone H2A (Lys9) (83041-1-RR) or 5 ug of Normal Rabbit IgG (98136-1-RR), and 20 µl of Protein A Magarose Beads. The immunoprecipitated DNA was quantified by real-time PCR.
WB Figures
WB analysis of NIH/3T3 using 83041-1-RR
Trichostatin A treated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 83041-1-RR (Acetyl-Histone H2A (Lys9) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with Beta Actin Monoclonal antibody (66009-1-Ig) as loading control.
IHC Figures
IHC staining of mouse lung using 83041-1-RR
Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 83041-1-RR (Acetyl-Histone H2A (Lys9) antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IHC staining of mouse lung using 83041-1-RR
Immunohistochemical analysis of paraffin-embedded mouse lung tissue slide using 83041-1-RR (Acetyl-Histone H2A (Lys9) antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IF/ICC Figures
IF Staining of HeLa using 83041-1-RR
Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Acetyl-Histone H2A (Lys9) antibody (83041-1-RR, Clone: 230349B1 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).
IF Staining of HeLa using 83041-1-RR
Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Acetyl-Histone H2A (Lys9) antibody (83041-1-RR, Clone: 230349B1 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).
IF Staining of HeLa using 83041-1-RR
Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Acetyl-Histone H2A (Lys9) antibody (83041-1-RR, Clone: 230349B1) at dilution of 1:150 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).
DOT BLOT Figures
Dot Blot experiment of peptide using 83041-1-RR
Dot blot analysis was used to confirm the specificity of Acetyl-Histone H2A (Lys9) antibody. Acetylated peptides were spotted onto NC and probed with antibody at 1 µg/ml.The amount of peptide (μg/mL) spotted is indicated next to each row.Column 1: H2AEK9Ac. Column 2: Unmodified H2AEK9. Column 3: H2AEK5Ac. Column 4: Unmodified H2AEK5. Column 5: H2AEK13Ac. Column 6: Unmodified H2AEK13. Column 7: H2AEK15Ac. Column 8: Unmodified H2AEK15. Column 9: Blank(PBS).
The species listed in Tested Reactivity are in-house verified and applicable species. For unlisted species, please refer to the homology analysis of the immunogen sequence and related species. For rabbit polyclonal antibodies, homology >70% is recommended. For mouse monoclonal antibodies and rabbit recombinant antibodies, homology >90% is recommended. Generally, the higher the homology, the greater the applicability. However, there will be certain differences in protein expression in different species, tissues or cells. Therefore, the homology analysis results are for reference only and do not serve as a guarantee.
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