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Acetylated-Lysine Recombinant Rabbit Monoclonal Antibody [PSH09-15] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751269

Acetylated-Lysine Recombinant Rabbit Monoclonal Antibody [PSH09-15] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IF-Tissue

  • ChIP

  • IP

Reactivity

  • Species independent

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

Acetylated-Lysine Recombinant Rabbit Monoclonal Antibody [PSH09-15] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic Acetylated lysine-containing peptide.

Species Reactivity

Species independent

Validated Applications

WB, IHC-P, IF-Tissue, ChIP, IP

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.

Conjugation

unconjugated

Clone Number

PSH09-15

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:20,000-1:100,000

  • IF-Tissue

  • 1:5,000-1:20,000

  • ChIP

  • Use 5 μg for 25 μg of chromatin.

  • IP

  • 1-2μg/sample

Target

Function

Acetylation of lysine, like phosphorylation of serine, threonine or tyrosine, is an important reversible modification controlling protein activity. The conserved amino-terminal domains of the four core histones (H2A, H2B, H3, and H4) contain lysines that are acetylated by histone acetyltransferases (HATs) and deacetylated by histone deacetylases (HDACs) . Signaling resulting in acetylation/deacetylation of histones, transcription factors, and other proteins affects a diverse array of cellular processes including chromatin structure and gene activity, cell growth, differentiation, and apoptosis. Recent proteomic surveys suggest that acetylation of lysine residues may be a widespread and important form of post-translational protein modification that affects thousands of proteins involved in control of cell cycle and metabolism, longevity, actin polymerization, and nuclear transport.

Background References

暂无

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Acetylated-Lysine on different lysates with Rabbit anti-Acetylated-Lysine antibody (<a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 1μM TSA for 18 hours cell lysate<br />Lane 3: NIH/3T3 cell lysate<br />Lane 4: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate<br />Lane 5: C6 cell lysate<br />Lane 6: C6 treated with 1μM TSA for 18 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Observed band size: Mutiple bands<br /><br />Exposure time: 42 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of Acetylated-Lysine on different lysates with Rabbit anti-Acetylated-Lysine antibody (HA751269) 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 (HA751269) 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.

  • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Acetylated-Lysine antibody (<a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a>) at 1/100,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a>) 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.

    Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Acetylated-Lysine antibody (HA751269) at 1/100,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA751269) 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.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Acetylated-Lysine antibody (<a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a>) at 1/20,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a>) 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.

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Acetylated-Lysine antibody (HA751269) at 1/20,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA751269) 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.

  • Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Acetylated-Lysine antibody (<a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a>) at 1/20,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a>) 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.

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Acetylated-Lysine antibody (HA751269) at 1/20,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA751269) 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.

  • Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Acetylated-Lysine antibody (<a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a>) at 1/100,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a>) 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.

    Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Acetylated-Lysine antibody (HA751269) at 1/100,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA751269) 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.

  • Indirect ELISA analysis of Acetylated-Lysine on different conjugations.

    Indirect ELISA analysis of Acetylated-Lysine on different conjugations.

  • Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Acetylated-Lysine (<a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a>) 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.

    Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Acetylated-Lysine (HA751269) 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.

  • Acetylated-Lysine was immunoprecipitated from 0.2 mg NIH/3T3 treated with 400nM TSA for 18 hours cell lysate with <a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Histone H3 (acetyl K9) (<a href="/products/HA722132" style="font-weight: bold;text-decoration: underline;">HA722132</a>) at 1/2,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (<a href="/products/M1208-2" style="font-weight: bold;text-decoration: underline;">M1208-2</a>) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: NIH/3T3 treated with 400nM TSA for 18 hours cell lysate (input)<br />Lane 2: <a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a> IP in NIH/3T3 treated with 400nM TSA for 18 hours cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA751269" style="font-weight: bold;text-decoration: underline;">HA751269</a> in NIH/3T3 treated with 400nM TSA for 18 hours cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 1 minute 57 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    Acetylated-Lysine was immunoprecipitated from 0.2 mg NIH/3T3 treated with 400nM TSA for 18 hours cell lysate with HA751269 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: HA751269 IP in NIH/3T3 treated with 400nM TSA for 18 hours cell lysate
    Lane 3: Rabbit IgG instead of HA751269 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

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