Human/Mouse/Rat Ezrin Antibody

Catalog #: AF7239 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF7239
AF7239-SP

Detection of Human, Mouse, and Rat Ezrin by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse/Rat Ezrin Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human, mouse, and rat Ezrin in Western blots and detects recombinant human Ezrin in direct ELISAs.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Ezrin
Lys438-Arg562
Accession # P15311

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.2 µg/mL

See below

Immunohistochemistry

0.5-15 µg/mL

Immersion fixed paraffin-embedded sections of human colon (colon stem cells differentiating into ascending colon) and human stomach

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.

Scientific Data

Western Blot

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Detection of Human, Mouse, and Rat Ezrin by Western Blot. Western blot shows lysates of human placenta tissue, mouse placenta tissue, and C6 rat glioma cell line. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human/Mouse/Rat Ezrin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7239) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (HAF016). A specific band was detected for Ezrin at approximately 81 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry

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Ezrin in Human Colon Stem Cells. Ezrin was detected in immersion fixed paraffin-embedded sections of human colon stem cells differentiating into ascending colon using Sheep Anti-Human/Mouse/Rat Ezrin Antigen Affinity-purified Polyclonal Antibody (AF7239) at 0.5 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; NL010). Chromogranin A was also detected using Mouse Anti-Human Chromogranin A Monoclonal Antibody (MAB90981) at 0.5 µg/mL overnight at 4 °C. Tissue was co-stained using the NorthernLights™ 637-conjugated Anti-Mouse IgG Secondary Antibody (red; NL008) and counterstained with DAPI (blue). Specific staining of Ezrin was localized to microtubules. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Immunohistochemistry

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Detection of Ezrin in Human Stomach Ezrin was detected in immersion fixed paraffin-embedded sections of human stomach using Sheep Anti-Human/Mouse/Rat Ezrin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7239) at 1 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Sheep IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC006). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surface of epithelial cells in gastric glands. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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: Ezrin

EZRIN (also Cytovillin, Villin2 and p81) is a founding member of the ERM family, Band 4.1 Superfamily of proteins. Although its predicted MW is 69 kDa, it runs anomalously at 77-82 kDa in SDS-PAGE. ERZIN is expressed by epithelial cells where it serves as a linker between the cell membrane and the actin cytoskeleton. Its presence is particularly strong in microvilli where it helps organize this structure. In addition, ERZIN also organizes microtubules in lymphocytes at or near the immunological synapse by interacting with Glg1. Human EZRIN is 585 amino acids (aa) in length. It contains a band 4.1 homology/FERM domain that binds CD44, ICAM-1, EBP50 and ERM family members (aa 1-295), a central alpha -helical region (aa 296-352), and a C-terminal ERM and actin-binding/FERM C domain (aa 353-586). EZRIN exists as either a monomer, or a homo/heterodimer. EZRIN is not constitutively active, but must be phosphorylated and unfolded to bind to cytoplasmic proteins. Over aa 438-562, human EZRIN shares 96% aa identity with mouse EZRIN.

Entrez Gene IDs

7430 (Human); 22350 (Mouse); 54319 (Rat)

Alternate Names

CVIL; CVL; Cytovillin 2; Cytovillin; DKFZp762H157; EZR; Ezrin; FLJ26216; p81; VIL2; VIL2MGC1584; villin 2 (ezrin); Villin 2; villin-2