Human beta-III Spectrin Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Human
  • Specificity
    Detects human beta-III Spectrin in direct ELISAs and Western blots.
  • Source
    Recombinant Monoclonal Rabbit IgG Clone # 1287A
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    Synthetic peptide containing the C-terminal region of beta-III Spectrin
    Accession # NP_008877.1
  • 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
    2 µg/mL
    See below
  • Immunohistochemistry
    8-25 µg/mL
    See below
  • Immunocytochemistry
    8-25 µg/mL
    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 beta-III Spectrin by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human beta-III Spectrin Monoclonal Antibody (Catalog # MAB9394) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for beta-III Spectrin at approximately 250 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
beta-III Spectrin in HeLa Human Cell Line. beta-III Spectrin was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Rabbit Anti-Human beta-III Spectrin Monoclonal Antibody (Catalog # MAB9394) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunohistochemistry
beta-III Spectrin in Human Brain. beta-III Spectrin was detected in immersion fixed paraffin-embedded sections of human brain (cerebellum) using Rabbit Anti-Human beta-III Spectrin Monoclonal Antibody (Catalog # MAB9394) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in Purkinje neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
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: beta-III Spectrin
Spectrin beta-III (SPTBN2) belongs to spectrins family of proteins which are essential to mechanical support/structural membrane integrity maintenance in erythrocytes, stabilizing cell-cell contacts, and localizing ion channels as well as cell adhesion molecules within specific subdomains of plasma membranes. Vertebrate spectrins have two alpha-subunits (alpha-I/alpha-II), four beta-subunits (beta-I-beta-IV) and a beta-H subunit creating diversity and specialization of function. Spectrin beta 3 is primarily expressed in nervous tissues with highest expression levels in the cerebellum, where it is found in Purkinje cell soma and dendrites. Spectrin beta-III interacts with EAAT4, the glutamate transporter predominately expressed in Purkinje cells, and stabilizes it at the plasma membrane leading to glutamate clearance from the synaptic cleft, and resulting in both modulation of glutamatergic neurotransmission as well as prevention of glutamate-mediated neurotoxicity.
  • Long Name:
    Spectrin Beta, Non-Erythrocytic 2
  • Entrez Gene IDs:
    6712 (Human); 29211 (Rat)
  • Alternate Names:
    beta-III Spectrin; glutamate transporter EAAT4-associated protein 41; GTRAP41; KIAA0302; SCA5; SCAR14; Spectrin beta 3; spectrin beta chain, brain 2; Spectrin beta III; spectrin, beta, non-erythrocytic 2; Spectrin, non-erythroid beta chain 2; spinocerebellar ataxia 5; SPTBN2
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-Universal

VCTS023
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VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
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VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
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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™ NL493-conjugated Antibody

Flow, IHC, ICC NL006 4
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Donkey Anti-Rabbit IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL005 2
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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 Antibody

WB AF008 1
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Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC003  
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Donkey Anti-Rabbit IgG (H+L) Antibody

D-301-C-ABS2
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