![Western Blot: Histone H2a [ac Lys5] Antibody [NBP2-59261] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H2AK5ac. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (kDa) is shown on the left](http://images.novusbio.com/fullsize/Histone-H2a-[ac-Lys5]-Antibody-Western-Blot-NBP2-59261-img0001.jpg "Western Blot: Histone H2a [ac Lys5] Antibody [NBP2-59261] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H2AK5ac. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (kDa) is shown on the left")
| Reactivity | HuSpecies Glossary |
| Applications | WB, DB, ELISA, ICC/IF, ChIP |
| Clonality | Polyclonal |
| Host | Rabbit |
| Conjugate | Unconjugated |
| Immunogen | The exact immunogen is propietary information. |
| Modification | ac Lys5 |
| Isotype | IgG |
| Clonality | Polyclonal |
| Host | Rabbit |
| Gene | H2AC20 |
| Purity | Peptide affinity purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
| Dilutions |
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| Storage | Store at -20C. Avoid freeze-thaw cycles. |
| Buffer | PBS |
| Preservative | 0.05% Sodium Azide and 0.05% ProClin 300 |
| Purity | Peptide affinity purified |
Secondary Antibodies |
Isotype Controls |
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| Gene Symbol | H2AC20 |
![Immunocytochemistry/Immunofluorescence: Histone H2a [ac Lys5] Antibody [NBP2-59261] - HeLa cells were stained with the antibody against H2AK5ac and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H2AK5ac antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa Fluor 488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.](http://images.novusbio.com/fullsize/Histone-H2a-[ac-Lys5]-Antibody-Immunofluorescence-NBP2-59261-img0004.jpg "Immunocytochemistry/Immunofluorescence: Histone H2a [ac Lys5] Antibody [NBP2-59261] - HeLa cells were stained with the antibody against H2AK5ac and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H2AK5ac antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa Fluor 488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.")
![Chromatin Immunoprecipitation: Histone H2a [ac Lys5] Antibody [NBP2-59261] - ChIP assays were performed using human HeLa cells, the antibody against H2AK5ac and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 0.5, 1, 2 and, 5 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the GAPDH and ACTB promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).](http://images.novusbio.com/fullsize/Histone-H2a-[ac-Lys5]-Antibody-Chromatin-Immunoprecipitation-NBP2-59261-img0003.jpg "Chromatin Immunoprecipitation: Histone H2a [ac Lys5] Antibody [NBP2-59261] - ChIP assays were performed using human HeLa cells, the antibody against H2AK5ac and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 0.5, 1, 2 and, 5 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the GAPDH and ACTB promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).")
![ELISA: Histone H2a [ac Lys5] Antibody [NBP2-59261] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H2AK5ac in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:25,000.](http://images.novusbio.com/fullsize/Histone-H2a-[ac-Lys5]-Antibody-ELISA-NBP2-59261-img0002.jpg "ELISA: Histone H2a [ac Lys5] Antibody [NBP2-59261] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H2AK5ac in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:25,000.")
![Dot Blot: Histone H2a [ac Lys5] Antibody [NBP2-59261] - To test the cross reactivity of the antibody against H2AK5ac, a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H2AK5. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure shows a high specificity of the antibody for the modification of interest.](http://images.novusbio.com/fullsize/Histone-H2a-[ac-Lys5]-Antibody-Dot-Blot-NBP2-59261-img0006.jpg "Dot Blot: Histone H2a [ac Lys5] Antibody [NBP2-59261] - To test the cross reactivity of the antibody against H2AK5ac, a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H2AK5. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure shows a high specificity of the antibody for the modification of interest.")
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![Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).](https://images.novusbio.com/images/Goat-anti-Rabbit-IgG-H+L-Secondary-Antibody-HRP-Western-Blot-NB7160-img0001.jpg "Western Blot: Goat anti-Rabbit IgG (H+L) Secondary Antibody [HRP] [NB7160] - Western blot showing vemurafenib treatment in BRAFV600E CRC cells inhibits fission mediator DRP1 with no significant effect on fusion proteins (Mfn1 & 2) using MFN-1 antibody (NBP1-51841) and corresponding secondary antibody, goat anti-rabbit IgG-HRP (NB7160). Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/33738242).")
![Flow Cytometry: Rabbit IgG Isotype Control [NBP2-24891] - Intracellular FACS analysis of mouse TLR6 polyclonal antibody (red), rabbit isotype control (green), RAW cells alone (shaded). Two micrograms of antibodies were used. Goat anti-rabbit FITC (Novus, 20302) was used as secondary.](https://images.novusbio.com/images/Rabbit--Mouse-IgG-Isotype-Control-Flow-Cytometry-NBP2-24891-img0001.jpg "Flow Cytometry: Rabbit IgG Isotype Control [NBP2-24891] - Intracellular FACS analysis of mouse TLR6 polyclonal antibody (red), rabbit isotype control (green), RAW cells alone (shaded). Two micrograms of antibodies were used. Goat anti-rabbit FITC (Novus, 20302) was used as secondary.")