Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- ELISA [2]
- Chromatin Immunoprecipitation [1]
- Other assay [1]
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Validation data
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- Product number
- 49-1022 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- H4K8ac Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- Conc. Not Determined
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Histone extracts of HeLa cells (15 µg) were analyzed by western blot using the anti-H4K8ac crude serum (Product # 49-1022), diluted 1:250 in TBS-Tween containing 5% milk. The position of the protein of interest is indicated with an arrow; the marker (in kDa) is shown on the right. Lane 2 shows the result of the western analysis with the crude serum; lane 1 shows the same analysis after incubation of the crude serum with 750 pmol blocking peptide for 1 hour at room temperature.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- To determine the titer, an ELISA was performed using the anti-H4K8ac crude serum (Product # 49-1022). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the crude serum was estimated to be 1:17, 500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- To determine the titer, an ELISA was performed using a serial dilution of the anti-human H4K8ac antibody (Product # 49-1022). 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 1:17,500.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- ChIP assays were performed using human osteosarcoma (U-2OS) cells, the anti-H4K8ac crude serum (Product # 49-1022), and optimized PCR primer sets for qPCR. ChIP was performed using 100,000 million cells treated with the deacetylase inhibitor ATRA. A titration of the crude serum consisting of 0.5, 1.5, and 5 µl per ChIP experiment was analyzed. Additionally, ChIP was performed after incubation of the crude serum with 5 nmol blocking peptide for 1 hour at room temperature. IgG (5 µg⁄IP) was used as negative IP control. qPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. This figure shows the recovery, expressed as a percentage of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- A dot blot analysis was performed to test the cross reactivity of the anti-H4K8ac crude serum (Product # 49-1022) with other modifications of histone H4 and H3 and with the unmodified histone H4. 100 to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. This figure shows a high specificity of the crude serum for the modification of interest.