Histone H3 Monoclonal antibody

Histone H3 Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

human, mouse, rat, pig, rabbit, canine, chicken, zebrafish, hamster, dog, wheat

Applications

WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Conjugate

Unconjugated

Publications

37

CloneNo.

1A2A3

Cat no : 68345-1-Ig

Synonyms

HIST1H3A, Histone-H3, HistoneH3, Histone H 3, 1A2A3

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Validation Data Gallery

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  WB analysis using 68345-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68345-1-Ig (Histone H3 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis using 68345-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 68345-1-Ig (HIST1H3A antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 68345-1-Ig

  IP result of anti-Histone H3 (IP:68345-1-Ig, 4ug; Detection:68345-1-Ig 1:2000) with HeLa cells lysate 1800 ug.

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  IHC staining of mouse testis using 68345-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 68345-1-Ig (HIST1H3A antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse testis using 68345-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse testis tissue slide using 68345-1-Ig (HIST1H3A antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of MCF-7 using 68345-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using Histone H3 antibody (68345-1-Ig, Clone: 1A2A3, labeled with CoraLite647-conjugated AffiniPure F(ab')2 Fragment Goat Anti-Mouse IgG (H+L),SA00014-10, magenta) at dilution of 1:400 and CoraLite Plus 488-conjugated ZO-1 antibody (CL488-21773, green), CHCHD6 antibody (66597-1-Ig, Clone: 2A11E9, labeled with FlexAble CoraLite Plus 555 Antibody Labeling Kit for Mouse IgG2a, KFA042, orange).

• 

  IF Staining of HeLa using 68345-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Histone H3 antibody (68345-1-Ig, Clone: 1A2A3 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), CL594-phalloidin (red).

• 

  IF Staining of A431 using 68345-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed A431 cells using Histone H3 antibody (68345-1-Ig, Clone: 1A2A3 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), CL594-phalloidin (red).

• 

  IF Staining of HeLa using 68345-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Histone H3 antibody (68345-1-Ig, Clone: 1A2A3 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), CL594-phalloidin (red).

• 

  IF Staining of MCF-7 using 68345-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using Histone H3 antibody (68345-1-Ig, Clone: 1A2A3 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), CL594-phalloidin (red).

• 

  IF Staining of HeLa using 68345-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Histone H3 antibody (68345-1-Ig, Clone: 1A2A3 ) at dilution of 1:1000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), Alpha Tubulin antibody (11224-1-AP, red).

• 

  FC experiment of HepG2 using 68345-1-Ig

  1X10^6 HepG2 cells were intracellularly stained with 0.4 ug Anti-Human Histone H3 (68345-1-Ig, Clone:1A2A3) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG2b Isotype Control (MPC-11) (65128-1-Ig, Clone: MPC-11) (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive WB detected in: LNCaP cells, HEK-293 cells, Jurkat cells, K-562 cells, HSC-T6 cells, NIH/3T3 cells, MDCK cells, CHO cells, HeLa cells, chicken brain tissue, zebrafish, wheat whole plant
• Positive IP detected in: HeLa cells
• Positive IHC detected in: mouse testis tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: MCF-7 cells, A431 cells, HeLa cells
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:500-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:500-1:2000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 36 publications below
• IF: See 2 publications below

Product Information

68345-1-Ig targets Histone H3 in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse, rat, pig, rabbit, canine, chicken, zebrafish, hamster, dog, wheat samples.

• Tested Reactivity: human, mouse, rat, pig, rabbit, canine, chicken, zebrafish, hamster, dog, wheat
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: histone cluster 1, H3a
• Observed Molecular Weight: 15 kDa
• GenBank Accession Number: BC066245
• Gene Symbol: HIST1H3A
• Gene ID (NCBI): 8350
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Histones are small, highly basic proteins that consist of a globular domain with unstructured N- and C-terminal tails protruding from the main structure. Histone H3 is one of the five main histones that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. In addition to their role in DNA compartmentalization, histones also play crucial roles in various biologic processes, including gene expression and regulation, DNA repair, chromatin condensation, cell cycle progression, chromosome segregation, and apoptosis. The ability of histones to regulate chromatin dynamics primarily originates from various posttranslational modifications carried out by histone-modifying enzymes.

Publications

Species	Application	Title
	WB	Nucleic Acids Res; The origin recognition complex requires chromatin tethering by a hypervariable intrinsically disordered region that is functionally conserved from sponge to man; Authors - Olubu A Adiji
mouse	WB	Redox Biol; Deficiency of S100 calcium binding protein A9 attenuates vascular dysfunction in aged mice; Authors - Boying Zhao
mouse	WB	Environ Pollut; Chronic exposure to polystyrene microplastics induced LHR reduction and decreased testosterone levels through NF-κB pathway; Authors - Haibo Jin
rat	WB	Int J Mol Sci; Ursolic Acid Alleviates Neuroinflammation after Intracerebral Hemorrhage by Mediating Microglial Pyroptosis via the NF-κB/NLRP3/GSDMD Pathway; Authors - Pan Lei
mouse	WB	Int Immunopharmacol; Azithromycin regulates Mettl3-mediated NF-κB pathway to enhance M2 polarization of RAW264.7 macrophages and attenuate LPS-triggered cytotoxicity of MLE-12 alveolar cells; Authors - Shuna Xu
human	WB	Curr Med Chem; Resveratrol Inhibits Hepatocellular Carcinoma Progression Through Regulating Exosome Secretion; Authors - Kun Tong