Anti-Histone H3 (acetyl K14) antibody [EP964Y] - BSA and Azide free

5 product images

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  ChIP - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - BSA and Azide free (ab203952)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade ab52946).
  

  Chromatin was prepared from HeLa cells (treated with 50 ng/ml nocodazole for 14 hours) according to the Abcam X-ChIP protocol. Cells were fixed with EGS (1.5 mM) for 30 minutes then formaldehyde (1%) for 10 minutes. The ChIP was performed with 25μg of chromatin, 5μg of Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade ab52946 (red), and 20μl of Protein A/G sepharose beads. No antibody was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (TaqMan approach).

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  Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - BSA and Azide free (ab203952)

  Immunocytochemistry/Immunofluorescence analysis of untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) cells and TSA (Trichostatin A) (500ng/ml, 4h) and treated HeLa cells labeling Histone H3 (acetyl K14) with purified Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade ab52946 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100, counterstained with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 AlexaFluor^®594 Goat anti-Mouse secondary 1:1000  (2ug/ml). An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077). Nuclei counterstained with DAPI (blue).

  Negative Control 1: Rabbit primary antibody and anti-mouse secondary antibody(Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120)

  Negative Control 2: Mouse primary antibody(Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) and anti-rabbit secondary antibody(Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade ab52946).

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  Western blot - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - BSA and Azide free (ab203952)

  This data was developed using Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade ab52946, the same antibody clone in a different buffer formulation. Different batches of Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade ab52946 were tested on C6 (Rat glial tumor glial cell) treated with Trichostatin A lysate at 1.2 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 15 kDa.

  All lanes: Western blot - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade (Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade ab52946)

  Predicted band size: 15 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - BSA and Azide free (ab203952)

  Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade ab52946 at 1/100 dilution staining Histone H3 (acetyl K14) in human uterus adenocarcinoma tissue by Immunohistochemistry, Paraffin embedded tissue.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade ab52946).

  Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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  Peptide Array - Anti-Histone H3 (acetyl K14) antibody [EP964Y] - BSA and Azide free (ab203952)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade ab52946).
  
  Anti-Histone H3 (acetyl K14) antibody [EP964Y] - ChIP Grade ab52946 at 0.01 μg/mL 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). A Goat Anti-Rabbit IgG, (H+L), Fluo 647nm conjugated at 1/50000 dilution was used as the secondary antibody. Washing buffer and time was protein Array Washing Buffer, 2*10mins and 2*5mins and blocking buffer was 5% BSA in TBST.
  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 the area under the curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
  
  Scanner details: Innoscan 710