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Recombinant Rabbit Monoclonal
Anti-Ubiquityl-Histone H2A (Lys119) Rabbit mAb
Catalog Number: PTM-1121
$ 320

Clone Number: JRMR-353

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB IHC-P ICC/IF IP ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H2AK119ub

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 H2AK119ub
UniProt ID

P04908

Immunogen Ubiquitinated human histone H2A (Lys119) peptide
MW (kDa) 23
Specificity Anti-Ubiquityl-Histone H2A (Lys119) Rabbit mAb detects histone H2A only when it is ubiquitinated at Lys119.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
IHC-P 1:50 - 1:100 Human, Mouse
ICC/IF 1:100 Human
ChIP 6 ug per 5X106 cells Human
IP 1:50 - 1:100 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

Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. Ubiquitination, a post-translational modification in which the ubiquitin is covalently attached to a substrate protein, occurs on histones H2A and H2B. Histone H2A is mono-ubiquitinated at Lys119 by the Polycomb Repressor Complex 1 (PRC1) and is critical for gene silencing, X chromosome inactivation, and cellular response to DNA damage. Histone H2A is also monoubiquitinated at Lys119 at sites of DNA damage.

Cellular location

Nucleus

Images
WB

Lysate: HeLa, N2a, BRL, mouse liver, mouse kidney
Protein loading amount: 20 μg
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000
Primary Ab incubation: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Exposure time: 8 seconds
Predicted band size: 23 kDa
Observed band size: 23 kDa
IHC-P

Tissue: Human hepatocellular carcinoma
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: 1:100
Primary Ab incubation: 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-1121.

Tissue: Mouse kidney
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: 1:100
Primary Ab incubation: 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-1121.
ICC/IF

Cell line: HeLa
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:100
Primary Ab incubation: 4°C overnight
Secondary Ab: Goat Anti-Rabbit IgG
Nuclear counter stain: DAPI (blue)
Counter stain: Tubulin (red)
Description: The green color represents the positive signal observed with PTM-1121.
ChIP

Cell type: HeLa
Cross-linking conditions: no cross-linking
Amount of chromatin per IP: 5×106 cells
Amount of Ab per IP: 6 ug
Beads type and amount per IP: 50 μL of Protein A MagBeads
Comment: The ChIP was performed with 6 μg of normal Rabbit IgG as a negative control. Real time quantitative PCR was performed on immunoprecipitated DNA using primers specific for the human GAPDH CDS region, LDHA, FOXO3a promoter, and TUBBP10. Data are presented as enrichment of each sample relative to the total amount of input chromatin at each amplicon.
IP

IP of HeLa cells extracts
IP Ab incubation: PTM- 1121, 1:100 dilution, 4°C overnight
WB Primary Ab incubation: PTM1121, room temperature 2h, 1: 1000 dilution
Secondary Ab: Anti-Rabbit IgG for IP (HRP)
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
1: 5% Input
2: IP with PTM-1121
3: IP with Rabbit monoclonal IgG Isotype
Observed MW: 23 kDa
Exposure time: 60 s

Research Use

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

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