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Recombinant Rabbit Monoclonal
Anti-Acetyllysine Rabbit mAb
Catalog Number: PTM-105RM
$ 320

Clone Number: PA-070-53

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB IHC-P IP ChIP

Reactivity: All

Synonyms: Kac

Product Size
100 μl
Quantity

Shipping: Ambient temperature

Order online or send purchase order to info@ptmbio.com

FAQ Technical Support Protocols

General Information
Isotype IgG
Conjugate Unconjugated
Synonyms Kac
UniProt ID

/

Immunogen Acetylated lysine peptides
MW (kDa) Multiple
Specificity Anti-Acetyllysine Rabbit mAb detects proteins with acetylated lysine residues. This pan antibody recognizes acetylated lysine independent of its surrounding sequences.
Product Usage Information
Applications Dilution Recommended Species
WB 1:1000 All
IHC-P 1:200 - 1:1000 Human, Mouse, Rat
ChIP 6 μg per 5x106 cells All
Properties
Purity Protein A purified
Constituents PBS, Glycerol, BSA
Storage Store at -20°C. Avoid freeze/thaw cycles.
Stability Stable for 12 months from date of receipt/reconstitution.
Target Information

Background

The reversible lysine acetylation of histones and non-histone proteins is a crucial post-translational modification that regulates diverse cellular processes, including chromatin dynamics, gene expression, cell cycle progression, apoptosis, differentiation, DNA replication, DNA repair, nuclear import, and neuronal repression. More than 20 acetyltransferases and 18 histone deacetylases (HDACs) have been identified, but the mechanisms governing substrate selection and site specificity of these enzymes remain unclear. Dysregulation of protein acetylation has been implicated in the development of cancers and other diseases, and HDACs have become likely targets for anti-cancer drugs.

Cellular location

/

Images
Dot Blot

Peptide amount: 4 ng, 20ng, 100 ng
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Exposure time: 30 seconds
The list of peptides used in the experiment is provided below.
Lane 1: Acetylated peptide library. Lane 2: Crotonylated peptide library.
Lane 3: Butyrylated peptide library. Lane 4: 2-Hydroxyisobutyrylated peptide library.
Lane 5: β-Hydroxybutyrylated peptide library. Lane 6: Unmodified peptide library.
WB

Lysate: Hela cells
Protein loading amount: 20 μg
Blocking buffer: 5% NFDM/TBST
Primary Ab: Lane 1, PTM-105RM pre-adsorbed with 3 μM synthetic acetylated lysine peptides; Lane 2, PTM-105RM pre-adsorbed with 3μM unmodified peptides.
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Exposure time: 30 seconds
Predicted band size: Multiple
Observed band size: Multiple

Lysates: 1: Hela; 2: NIH-3T3; 3: BRL; 4: Mouse liver; 5: Mouse kidney
Protein loading amount: 20 μg
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Exposure time: 60 seconds
Predicted band size: Multiple
Observed band size: Multiple

Lysates: (-): HeLa cells, (+): HeLa cells treated with 30 mM sodium butyrate for 4 hours
Protein loading amount: 20 μg
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Exposure time: 60+ seconds
Predicted band size: Multiple
Observed band size: Multiple

Lysates: (-): HepG2 cells, (+): HepG cells treated with 5 mM sodium butyrate for 24 hours
Protein loading amount: 20 μg
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Exposure time: 60+ seconds
Predicted band size: Multiple
Observed band size: Multiple
IHC-P

Tissue: Human liver
Section type: Formalin fixed & paraffin-embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter stain: Hematoxylin (blue)
Description: The brown color represents the positive signal observed with PTM-105RM.

Tissue: Mouse cerebrum
Section type: Formalin fixed & paraffin-embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter stain: Hematoxylin (blue)
Description: The brown color represents the positive signal observed with PTM-105RM.

Tissue: Rat liver
Section type: Formalin fixed & paraffin-embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0
Primary Ab dilution: 1:1000
Primary Ab incubation condition: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter stain: Hematoxylin (blue)
Description: The brown color represents the positive signal observed with PTM-105RM.
IP

IP of MCF-7 cells extracts
Lane 1: 5% Input
Lane 2: IP with Rabbit Monoclonal IgG Isotype
Lane 3: IP with PTM-105RM
IP Ab incubation condition: 4°C overnight, 1:50 dilution
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
WB Primary Ab incubation condition: PTM-113 H3K14ac pAb, 2 hours at room temperature, 1:2000 dilution
Secondary Ab: Goat Anti-Rabbit IgG pAb (HRP Conjugate)
Exposure time: 30 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa
ChIP

Sample: Hela cells treated with 5 mM sodium butyrate for 24 hours
Cross-linking conditions: No cross-linking
Amount of chromatin per IP: 5×106 cells
Amount of Ab per IP: 6 μg
Beads type and amount per IP: 50 μL of Protein A MagBeads
Description: Chromatin immunoprecipitations were performed with 6 μg of normal rabbit IgG as a negative control. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH-CDS, RPL30, LDHA promoter, LDHA CDS, FOXO3a promoter, FOXO3a-downstream, RAB20, and TUBBP10 regions. The data are presented as enrichment of each sample relative to total amount of input at each amplicon.

Research Use

For research use only, not for use in diagnostic procedures.

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    Authors Yu Zhang, et al.
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