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Recombinant Rabbit Monoclonal
Anti-Ubiquityl-Histone H2A (Lys119) Rabbit mAb
Catalog Number: PTM-1121
$ 330

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/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 condition: 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 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-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 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-1121.
ICC/IF

Cell line: HeLa
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:100
Primary Ab incubation condition: 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 total amount of input chromatin at each amplicon.
IP

IP of HeLa cells extracts
IP Ab incubation condition: PTM- 1121, 4°C overnight, 1:100 dilution
WB primary Ab incubation condition: 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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