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Recombinant Rabbit Monoclonal
Anti-Histone H3 Rabbit mAb
Catalog Number: PTM-1001RM
References (31)
$ 125

Clone Number: PAPTM-686-8

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB IHC-P ICC/IF FC ChIP

Reactivity: Human, Mouse, Rat

Synonyms: Histone H3.1, Histone H3/a, H3C1, H3FA, HIST1H3A, Histone H3/b, Histone H3/c, Histone H3/d

Product Size
100μl
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 Histone H3.1, Histone H3/a, H3C1, H3FA, HIST1H3A, Histone H3/b, Histone H3/c, Histone H3/d
UniProt ID

P68431

Immunogen Synthetic peptide corresponding to the N-terminus of human histone H3 protein
MW (kDa) 15
Specificity Anti-Histone H3 Rabbit mAb detects endogenous levels of total histone H3 protein. It does not cross-react with other histone or non-histone proteins.
Product Usage Information
Applications Dilution Recommended Species
WB 1:500 - 1:1000 Human, Mouse, Rat
IHC-P 1:100 - 1:500 Human, Mouse, Rat
ICC/IF 1:50 - 1:200 Human
FC 1:50 - 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

Histones are a family of basic proteins that form the core of the nucleosome – the fundamental structural unit of chromatin.  In a single nucleosome, core histone proteins H2A, H2B, H3, and H4 form an octamer around which the DNA is tightly wrapped. Histone proteins not only serve to compact chromosomal DNA but also play vital roles in the dynamic and long-term regulation of genes by a wide variety of post-translational modifications (PTMs). These PTMs 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. Among the histone members, histone H3 is the most extensively modified and its amino-terminal tail structure has been shown to be especially critical for eukaryotic cells.

Cellular location

Nucleus

Images
WB

Lysates: 1: HeLa, 2: NIH-3T3, 3: BRL, 4: Rat kidney, 5: Mouse kidney, 6: Rat brain, 7: Mouse brain
Protein loading amount: 20 μg
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: 30 seconds
Predicted band size: 15 kDa
Observed band size: 15 kDa
IHC-P

Tissue: Human prostate hyperplasia
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: 500
Primary Ab incubation: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter strain: Hematoxylin (blue)
Description: The brown color represents the positive signal observed with PTM-1001RM.

Tissue: Mouse colon
Section type: fixed & paraffin-embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0
Primary Ab dilution: 1: 500
Primary Ab incubation: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter strain: Hematoxylin (blue)
Description: The brown color represents the positive signal observed with PTM-1001RM.

Tissue: Rat colon
Section type: fixed & paraffin-embedded section
Retrieval method: High temperature and high pressure
Retrieval buffer: Tris/EDTA buffer, pH 9.0
Primary Ab dilution: 1: 500
Primary Ab incubation: 1 hour at room temperature
Secondary Ab: Anti-Rabbit and Mouse Polymer HRP (Ready to Use)
Counter strain: Hematoxylin (blue)
Description: The brown color represents the positive signal observed with PTM-1001RM.
ICC/IF

Cell line: HeLa
Fixative: 100% Ice-cold methanol
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:200
Primary 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-1001RM.
FC

Cell line: HeLa
Fixative: 4% Paraformaldehyde
Permeabilization: 90% methanol
Primary Ab dilution: 1:100
Secondary Ab: Goat Anti-Rabbit IgG
Unlabeled control: Cells without incubation with primary and secondary antibodies (black line).
Isotype control: Rabbit mAb IgG Isotype Control (blue line).
Description: Line red is the positive signal for PTM-1001RM.
ChIP

Cell type: HeLa
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, 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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    Authors Hang Yin, et al.
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    Journal iScience
    Authors Jianpeng Yu, et al.
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