Human HIST3H3 Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Human
  • Specificity
    Detects human HIST3H3 in Western blots.
  • Source
    Recombinant Monoclonal Rabbit IgG Clone # 2128A
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Human HIST3H3 synthetic peptide
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    See below
  • Immunohistochemistry
    3-25 µg/mL
    See below
  • Immunocytochemistry
    0.3-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human HIST3H3 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with Sodium (Na) Butyrate. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human HIST3H3 Monoclonal Antibody (Catalog # MAB9448) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for HIST3H3 at approximately 16 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
HIST3H3 in HeLa Human Cell Line. HIST3H3 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Rabbit Anti-Human HIST3H3 Monoclonal Antibody (Catalog # MAB9448) at 0.3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (yellow; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cell nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunohistochemistry
HIST3H3 in Human Testis. HIST3H3 was detected in immersion fixed paraffin-embedded sections of human testis using Rabbit Anti-Human HIST3H3 Monoclonal Antibody (Catalog # MAB9448) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Intracellular Staining by Flow Cytometry
Detection of HIST3H3 in HeLa cells by Flow Cytometry. HeLa human cell line was stained with Rabbit anti-Human HIST3H3 Monoclonal Antibody (Catalog # MAB9448, filled histogram) or Normal Rabbit IgG control (Catalog # MAB1050, open histogram) followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012).
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: HIST3H3
Histones are alkaline proteins which organize nuclear DNA into nucleosomes. Histone H3 is one of four core histones, which exist as dimers to form an octameric histone core. Epigenetic modification of the exposed histone tails by methylation or acetylation is known to regulate transcription in various ways and is known to affect the etiology of a variety of cancers and developmental defects.  Methylation by methyltranserases mediates transcriptional activation (SET1, Chd1), elongation (MLL, WDR5) or silencing (SET7/9, JMJD2A). Expression of H3K4 methyltransferases have been associated with solid tumors such as prostate, ovary, squamous cell carcinoma, hepatocellular carcinoma and cancers of the blood, including T-cell acute lymphoblastic leukemia, acute myeloid leukemia, acute lymphoblastic leukemia, and others.
  • Long Name:
    Histone Cluster 3 H3
  • Entrez Gene IDs:
    8290 (Human)
  • Alternate Names:
    H3 histone family, member T; H3.4; H3/G; H3/gH3.4; H3/T; H3FT; H3FTMGC126888; H3t; H3tMGC126886; HIST3H3; Histone 3, H3; histone cluster 3, H3; histone H3.1t
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit

CTS005 3
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Anti-Rabbit HRP-AEC Cell & Tissue Staining Kit

CTS006 3
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Immunohistochemistry Reagents
Description Application Cat# Citations Images  

Antigen Retrieval Reagent-Universal

CTS015 10
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Antigen Retrieval Reagent-Basic

CTS013 4
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Mounting Medium

CTS011 3
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Antigen Retrieval Reagent Sampler (50 mL each)

CTS016
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Antigen Retrieval Reagent-Acidic

CTS014
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NorthernLights Guard Mounting Media

NL996
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VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
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VisUCyte Antigen Retrieval Reagent-Universal

VCTS023
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VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
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Isotype Controls
Description Application Cat# Citations Images  

Normal Rabbit IgG Control

Ctrl AB-105-C 40  
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Normal Rabbit IgG Control

Ctrl, CyTOF-ready MAB1050  
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Secondary Antibodies
Description Application Cat# Citations Images  

Rabbit IgG HRP-conjugated Antibody

WB HAF008 18  
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Donkey Anti-Rabbit IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL004 10
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Rabbit IgG PE-conjugated Antibody

Flow F0110 2  
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Rabbit IgG APC-conjugated Antibody

Flow F0111 1  
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Donkey Anti-Rabbit IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL005 2
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Donkey Anti-Rabbit IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL006 4
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Rabbit IgG Fluorescein-conjugated Antibody

Flow F0112 5  
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Goat Anti-Rabbit IgG Biotinylated Antibody

WB BAF008 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC003  
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Rabbit IgG Antibody

WB AF008 1
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Donkey Anti-Rabbit IgG (H+L) Antibody

D-301-C-ABS2
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