Anti-Histone H4 (unmodified K20) antibody [EPR22116] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (unmodified K20) antibody [EPR22116] - BSA and Azide free (AB236241)

Immunohistochemical analysis of paraffin-embedded human bladder cancer tissue labeling Histone H4K20 with ab227804 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Sporadic nuclear staining in human bladder cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227804).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (unmodified K20) antibody [EPR22116] - BSA and Azide free (AB236241)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Histone H4K20 with ab227804 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Sporadic nuclear staining in human colon tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227804).

• ChIP

Supplier Data

ChIP - Anti-Histone H4 (unmodified K20) antibody [EPR22116] - BSA and Azide free (AB236241)

Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25 μg of chromatin, 2 μg of ab227804 (red), and 20 μl of protein A/G sepharose beads slurry (10 μl of sepharose A beads + 10 μl of sepharose G beads). 2 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (SYBR approach).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227804).

• IP

Unknown

Immunoprecipitation - Anti-Histone H4 (unmodified K20) antibody [EPR22116] - BSA and Azide free (AB236241)

Histone H4 (unmodified K20) was immunoprecipitated from 0.35 mg of SET8 depleted U-2 OS (human bone osteosarcoma epithelial cell line) whole cell lysate with ab227804 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227804 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : SET8 depleted U-2 OS whole cell lysate 10 μg (Input).

Lane 2 : ab227804 IP in SET8 depleted U-2 OS whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab227804 in SET8 depleted U-2 OS whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 30 seconds.

Depletion of Set8, a histone methyltransferase, delays S-phase progression and impairs DNA replication (PMID : 18166648). It also prevents the monomethylation of H4K20, which is normally mediated by Set8 (PMID : 18166648).

This lysate was kindly provided by our collaborator Dr. Anja Groth, Biotech Research and Innovation Centre (BRIC). University of Copenhagen.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227804).

All lanes:

Immunoprecipitation - Anti-Histone H4 (unmodified K20) antibody [EPR22116] - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h4-unmodified-k20-antibody-epr22116-chip-grade-ab227804'>ab227804</a>)

Predicted band size: 11 kDa

Observed band size: 12 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (unmodified K20) antibody [EPR22116] - BSA and Azide free (AB236241)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Histone H4K20 with ab227804 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in mouse spleen is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227804).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H4 (unmodified K20) antibody [EPR22116] - BSA and Azide free (AB236241)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling Histone H4K20 with ab227804 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Sporadic nuclear staining in rat spleen is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab227804).

• PepArr

Lab

Peptide Array - Anti-Histone H4 (unmodified K20) antibody [EPR22116] - BSA and Azide free (AB236241)

ab227804 was tested in Peptide array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate). Circle area represents affinity between the antibody and a peptide : all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.

The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, is available under the Product Protocol section.