Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [1]
- ELISA [1]
- Immunocytochemistry [1]
- Other assay [2]
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Validation data
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- Product number
- TA347152 - Provider product page
- Provider
- OriGene
- Product name
- Mouse Monoclonal H3K4me3 Antibody
- Antibody type
- Monoclonal
- Description
- Mouse Monoclonal H3K4me3 Antibody
- Host
- Mouse
- Conjugate
- Unconjugated
- Epitope
- HIST1H3A
- Isotype
- IgG
- Antibody clone number
- NULL
- Vial size
- 50 µg
- Concentration
- 1 ug/ul
Submitted references Early-Life Social Isolation-Induced Depressive-Like Behavior in Rats Results in Microglial Activation and Neuronal Histone Methylation that Are Mitigated by Minocycline.
Wang HT, Huang FL, Hu ZL, Zhang WJ, Qiao XQ, Huang YQ, Dai RP, Li F, Li CQ
Neurotoxicity research 2017 May;31(4):505-520
Neurotoxicity research 2017 May;31(4):505-520
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Supportive validation
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- WB using the antibody against H3K4me3 diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
- Validation comment
- WB
Supportive validation
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- Cross reactivity of the antibody against H3K4me3 To test the specificity an ELISA was performed using a serial dilution of the antibody against H3K4me3. The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Image shows a high specificity of the antibody for the modification of interest.
- Validation comment
- ELISA
Supportive validation
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- HeLa cells were stained with the antibody against H3K4me3 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
- Validation comment
- IF
Supportive validation
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 ug H3K4me3 ab. The IP'd DNA was analysed on an Illumina Genome Analyzer. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Image shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.
- Validation comment
- Assay
- Submitted by
- OriGene (provider)
- Main image
- Experimental details
- ChIP assays using HeLa cells: ChIP-seq kit, using sheared chromatin from 1 million cells. A titration of 1, 2, 5 and 10 ug ab was used (2ug/IP IgG as negative control). qPCR primers were specific to the promoter of constitutively expressed GAPDH and c-fos genes as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat 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): results in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.
- Validation comment
- Assay