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Recombinant Rabbit Monoclonal
Anti-Acetyl-Histone H2A (Lys9) Rabbit mAb
Catalog Number: PTM-194
$ 320

Clone Number: 34-H4L2

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB IP ChIP

Reactivity: Human, Mouse

Synonyms: H2AK9ac

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 H2AK9ac
UniProt ID

P04908

Immunogen Acetylated human histone H2A (Lys9) peptide
MW (kDa) 14
Specificity Anti-Acetyl-Histone H2A (Lys9) Rabbit mAb detects histone H2A only when it is acetylated at Lys9.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:1000 Human, Mouse
IP 1:25 - 1:100 Human
ChIP 6 μg per 5x106 cells Human
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

Histone post-translational modifications (PTMs), known as the “histone code”, are key mechanisms of epigenetics that modulate chromatin structures. The PTMs on histone including acetylation, methylation, phosphorylation, and novel acylations directly affect the accessibility of chromatin to transcription factors and other epigenetic regulators, altering genome stability and gene transcription. Histone acetylation, tightly controlled by the opposing action of histone acetyltransferases (HATs) and histone deacetylases (HDACs), occurs primarily at lysine residues on the N-terminal tails of histones H2A (Lys5, 9, and 15), H2B (Lys5, 12, 15, 16, and 20), H3 (Lys4, 9, 14, 18, 23, 27, and 36), and H4 (Lys5, 8, 12, 16, and 20), and plays vital roles in the regulation of gene expression, DNA damage repair, chromatin dynamics, etc.

Cellular location

Nucleus

Images
Dot Blot

Peptide amount: 1 ng, 4 ng, 16 ng, 64 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: 60 seconds
The list of peptides used in the experiment is provided below.
Lane 1: H2AK9 acetyl. Dot 2: H2AK9 2-hydroxyisobutyryl.
Dot 3: H2AK9 β-hydroxybutyryl. Dot 4: H2AK13 acetyl.
Dot 5: H2AK9 unmodified.
WB

Lysates: (-) MCF-7 cells; (+) MCF-7 cells + sodium butyrate (50 mM, 24 hours) + trichostatin A (1.65 μM, 4 hours)
Protein loading amount: 20 μg
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:1000
Primary Ab incubation: 4℃ overnight
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Exposure time: 60 seconds
Predicted band size: 14 kDa
Observed band size: 14 kDa

Lysates: (-) NIH/3T3 cells; (+) NIH/3T3 cells treated with 1 μM trichostatin A for 18 hours
Protein loading amount: 20 μg
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:1000
Primary Ab incubation: 4℃ overnight
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Exposure time: 60 seconds
Predicted band size: 14 kDa
Observed band size: 14 kDa
IP

IP of whole cell lysates from MCF-7 + sodium butyrate (50 mM, 24 hours) + trichostatin A (1.65 μM, 4 hours)
Lane 1: 5% Input
Lane 2: IP with PTM-194
Lane 3: IP with Rabbit mAb IgG Isotype Control
IP Ab incubation: 1:100 dilution, 4°C overnight
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
WB Primary Ab incubation: PTM-194, 1:1000 dilution, 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG for IP (HRP Conjugate)
Exposure time: 60 seconds
Predicted band size: 14 kDa
Observed band size: 14 kDa
ChIP

Sample: MCF-7 + sodium butyrate (50 mM, 24 hours) + trichostatin A (1.65 μM, 4 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 CDS, FOXO3a promoter, FOXO3a downstream, RAB20, and TUBBP10 regions. The data are presented as enrichment of each sample relative to the total amount of input chromatin at each amplicon.

Research Use

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

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