Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- ELISA [1]
- Other assay [2]
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Validation data
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- Product number
- TA347215 - Provider product page
- Provider
- OriGene
- Product name
- Rabbit Polyclonal H4K20me1 Antibody
- Antibody type
- Polyclonal
- Description
- Rabbit Polyclonal H4K20me1 Antibody
- Host
- Rabbit
- Conjugate
- Unconjugated
- Epitope
- HIST4H4
- Isotype
- IgG
- Antibody clone number
- NULL
- Vial size
- 50 µg
- Concentration
- 2 ?g/?l
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Supportive validation
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- WB was performed on whole cell extracts (25 ug, lane 1) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 2, 3, 4 and 5, respectively) using the antibody against H4K20me1. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right.
- Validation comment
- WB
Supportive validation
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- Determination of the titer To determine the titer, an ELISA was performed using a serial dilution of the antibody against H4K20me1 in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:51,100.
- Validation comment
- ELISA
Supportive validation
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- ChIP assays using HeLa cells (sheared chromatin from 1 million cells). Titration of 1, 2, 5, and 10ug antibody per ChIP was analysed. IgG (2 ug/IP) was used as negative control. qPCR primers were for the coding region of active GAPDH and ACTB genes as positive controls, and for GAPDH promoter and a region located 1 kb upstream of GAPDH promoter as negative controls. Image shows the recovery, expressed as a % of input (the relative amount of IP'd DNA compared to input DNA after qPCR).
- Validation comment
- Assay
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- A Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Image shows a high specificity of the antibody for the modification of interest.
- Validation comment
- DB