Diagenode

H3pan monoclonal antibody 1B1B2 - Classic (sample size)

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Catalog Number
Format
Price
C15200011-10
10 μg
$80.00
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Monoclonal antibody raised in mouse against histone H3, using a KLH-conjugated synthetic peptide containing an unmodified sequence from the C-terminus of the protein. This antibody can be used as a loading control in both ChIP and WB experiments.

Lot001
Concentration0.5 µg/µl
Species reactivityHuman, mouse, maize, tomato, poplar
TypeMonoclonal
PurityProtein A purified
HostMouse
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP 1-2 μg/ChIP Fig 1
Western Blotting 1:1,000 Fig 2, 3
Immunofluorescence 1:500 Fig 4

* Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.

  • Validation Data

    ChIP

    Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3
    ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3 (cat. No. C15200011) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (cat. No. AB-Auto02-A100), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    Western Blot

    Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against H3
    Western blot was performed on whole cell extracts from HeLa cells (25 μg, lane 1) and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 2, 3, 4 and 5, respectively) using the Diagenode monoclonal antibody against H3 (Cat. No. C15200011). The antibody was diluted 1:1,000 in TBSTween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.

    ChIP

    Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H3
    Western blot was performed on whole cell extracts (30 μg) from different celltypes (lane 1: HeLa, lane 2: K562, lane 3: MCF7, lane 4: U2OS, lane 5: HepG2, lane 6: Jurkat, lane 7: NIH3T3, lane 8: E14Tg2a mouse ES cells) using the Diagenode monoclonal antibody against H3 (Cat. No. C15200011). 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 of interest is indicated on the right.

    Immunofluorescence

    Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against H3
    HeLa cells were stained with the Diagenode monoclonal antibody against H3 (Cat. No. C15200011) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labeled with the H3 antibody (middle) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right

  • Loading Control

    H3pan monoclonal antibody can be used as a loading control for nuclear samples to compare the protein expression level between different samples.

    Why do you need a loading control?

    • To verify that equal sample amount was loaded on the gel
    • To confirm that the electrotransfer was done equally across the lines
    • To check that the signal detection after antibody incubation is homogenous across the lines

    What is the loading control?

    The loading control is an antibody against a protein which is highly expressed in certain types of samples. The level of protein stays constant in different conditions and in different organisms. The molecular weights of the protein of interest and loading control are different, allowing differentiation of both proteins as two different bands.

  • Applications
    ChIP-qPCR (ab)
    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
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
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    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
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    Datasheet H3pan C15200011 DATASHEET
    Monoclonal antibody raised in mouse against histone H3, using a KLH-conjugated synthetic peptide ...
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  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: H3pan monoclonal antibody 1B1B2 - Classic (sample size) (Diagenode Cat# C15200011-10 Lot# 001). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

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