Diagenode

H4R3me2(sym) polyclonal antibody - Classic

RFX5-polyclonal-antibody-diagenode
Catalog Number
Format
Price
C15410308
50 μg
$295.00
  Bulk order

Polyclonal antibody raised in rabbit against the region of histone H4 containing the symmetrically dimethylated arginine 3 (H4R3me2(sym)), using a KLH-conjugated synthetic peptide. The antibody also recognizes H2AR3me2(sym).

LotA2274P
Concentration0.6 µg/µl
Species reactivityHuman
TypePolyclonal
PurityAffinity purified
HostRabbit
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ELISA 1:500 Figure 1
Dot Blotting 1:5,000 Figure 2
Western Blotting 1:1,000 Figure 3
Immunofluorescence 1:500 Figure 4
  • Validation Data

    ELISA

    Figure 1. Determination of the antibody titer
    To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4R3me2(sym) (cat. No. 15410308) 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 1), the titer of the antibody was estimated to be 1:11,750.

    Cross reactivity

    Figure 2. Cross reactivity tests using the Diagenode antibody directed against H4R3me2(sym)
    To test the cross reactivity of the Diagenode antibody against H4R3me2(sym) (cat. No. 15410308), a Dot Blot analysis was performed with peptides containing other histone arginine methylations and the unmodified H4R3. 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 2 shows the antibody is specific for the H4R3 symmetric dimethylation and also recognizes the H2AR3 symmetric dimethylation.

    Western blot

    Figure 3. Western blot analysis using the Diagenode antibody directed against H4R3me2(sym)
    Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4R3me2(sym) (cat. No. C15410308). The antibody was 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.

    Immunofluorescence

    Figure 4. Immunofluorescence using the Diagenode antibody directed against H4R3me2(sym)
    HeLa cells were stained with the Diagenode antibody against H4R3me2(sym) (cat. C15410308) 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 H4R3me2(sym) antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

  • Applications
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    DB
    Dot blotting Read more
    WB
    Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more
    IF
    Immunofluorescence: Diagenode offers huge selection of highly sensitive antibodies validated in IF. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 HeLa cells transfected with a Cas9 expression vector (... Read more
  • Documents
    Datasheet H4R3me2 C15410308 DATASHEET
    Datasheet description
    Download
  • Publications

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