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Anti-Acetyl-Histone H2B (Lys11) Mouse mAb
Catalog Number: PTM-176
$ 370

Clone Number: /

Host: Mouse Clonality: Monoclonal

Applications： WB IHC-P ICC/IF IP ChIP

Reactivity: Human, Mouse, Rat

Synonyms: H2BK11ac

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 H2BK11ac
UniProt ID

P62807

Immunogen Acetylated human histone H2B (Lys11) peptide
MW (kDa) 14
Specificity Anti-Acetyl-Histone H2B (Lys11) Mouse mAb detects histone H2B only when it is acetylated at Lys11.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:2000 Human, Mouse, Rat
IHC-P 1:200 - 1:1000 Human, Mouse, Rat
ICC/IF 1:50 Human
IP 1:25 - 1:100 Human
ChIP 6 μg per 5x106 cells Human
Properties
Purity Protein G and immunogen affinity 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
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:1000
Primary Ab incubation: 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: H2BK11 acetyl. Lane 2: H2BK5 acetyl.
Lane 3: H2BK5 butyryl. Lane 4: H2BK5 succinyl.
Lane 5: H2BK5 crotonyl. Lane 6: H2BK5 2-hydroxyisobutyryl.
Lane 7: H2BK5 β-hydroxybutyryl. Lane 8: H2BK11 butyryl.
Lane 9: H2BK11 crotonyl. Lane 10: H2BK11 β-hydroxybutyryl.
Lane 11: H2BK12 acetyl. Lane 12: H2BK12 butyryl.
Lane 13: H2BK12 crotonyl. Lane 14: H2BK12 2-hydroxyisobutyryl.
Lane 15: H2BK12 β-hydroxybutyryl. Lane 16: H2BK11 unmodified.
WB

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: 1:2000
Primary Ab incubation: 2 hours at room temperature
Secondary Ab: Goat Anti-Mouse IgG H&L pAb (HRP Conjugate)
Exposure time: 60 seconds
Predicted band size: 14 kDa
Observed band size: 14 kDa

Lysates: Mouse liver, mouse spleen
Protein loading amount: 20 μg
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:500
Primary Ab incubation: 2 hours at room temperature
Secondary Ab: Goat Anti-Mouse IgG H&L pAb (HRP Conjugate)
Exposure time: 60 seconds
Predicted band size: 14 kDa
Observed band size: 14 kDa
IHC-P

Tissue: Human testis
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: 1:200
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-176.

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: 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-176.

Tissue: Rat spleen
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: 1:200
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-176.
ICC/IF

Samples: (A) HeLa cells; (B) HeLa cells treated with 30 mM sodium butyrate for 4 hours
Fixative: 4% Paraformaldehyde
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:50
Primary Ab incubation: 4°C overnight
Secondary Ab: Goat Anti-Mouse IgG
Nuclear counter stain: DAPI (blue)
Counter stain: Tubulin (red)
Description: The green color represents the positive signal observed with PTM-176.
IP

IP of whole cell lysates from HeLa cells treated with 30 mM sodium butyrate for 4 hours
Lane 1: 5% Input
Lane 2: IP with PTM-176, 1:25 dilution
Lane 3: IP with PTM-176, 1:100 dilution
IP Ab incubation: 1:25 and 1:100 dilutions, 4°C overnight
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
WB Primary Ab incubation: PTM-176, 1:2000 dilution, 2 hours at room temperature
Secondary Ab: Goat Anti-Mouse IgG for IP (HRP Conjugate)
Exposure time: 60 seconds
Predicted band size: 14 kDa
Observed band size: 14 kDa
ChIP

Sample: HeLa cells
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 G MagBeads
Description: Chromatin immunoprecipitations were performed with 6 μg of normal mouse IgG as a negative control. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human 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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