Acetylated-Lysine Recombinant Rabbit Monoclonal Antibody [PSH09-15]
cat.: HA723073
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Species independent
Applications: WB, IHC-P, IF-Tissue, ChIP, IP
Clonality: Monoclonal
Clone number: PSH09-15
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Isotype: IgG
Immunogen: Synthetic Acetylated lysine-containing peptide.
Positive control: HeLa cell lysate, HeLa treated with 1μM TSA for 18 hours cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 400nM TSA for 18 hours cell lysate, C6 cell lysate, C6 treated with 1μM TSA for 18 hours cell lysate, human breast cancer tissue, human colon cancer tissue, mouse liver tissue, rat liver tissue.
Recommended Dilutions:
  WB
  IHC-P
  IF-Tissue
  ChIP
  IP
1:2,000
1:20,000-1:100,000
1:5,000-1:20,000
Use 5 μg for 25 μg of chromatin.
1-2μg/sample
Images
HA723073_1.jpg Fig1: Western blot analysis of Acetylated-Lysine on different lysates with Rabbit anti-Acetylated-Lysine antibody (HA723073) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 1μM TSA for 18 hours cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate
Lane 5: C6 cell lysate
Lane 6: C6 treated with 1μM TSA for 18 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Observed band size: Mutiple bands

Exposure time: 42 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723073) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA723073_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Acetylated-Lysine antibody (HA723073) at 1/100,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723073) at 1/100,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723073_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Acetylated-Lysine antibody (HA723073) at 1/20,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723073) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723073_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Acetylated-Lysine antibody (HA723073) at 1/20,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723073) at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723073_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Acetylated-Lysine antibody (HA723073) at 1/100,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723073) at 1/100,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723073_6.jpg Fig6: Indirect ELISA analysis of Acetylated-Lysine on different conjugations.
HA723073_7.jpg Fig7: Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Acetylated-Lysine (HA723073) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
HA723073_8.jpg Fig8: Acetylated-Lysine was immunoprecipitated from 0.2 mg NIH/3T3 treated with 400nM TSA for 18 hours cell lysate with HA723073 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Histone H3 (acetyl K9) (HA722132) at 1/2,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate (input)
Lane 2: HA723073 IP in NIH/3T3 treated with 400nM TSA for 18 hours cell lysate
Lane 3: Rabbit IgG instead of HA723073 in NIH/3T3 treated with 400nM TSA for 18 hours cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 1 minute 57 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.