Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and A431 human epithelial carcinoma cell line. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human Eps15 Monoclonal Antibody (Catalog # MAB8480) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Eps15 at approximately 140 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Eps15 in HeLa Human Cell Line.

Eps15 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Rabbit Anti-Human Eps15 Monoclonal Antibody (Catalog # MAB8480) at 1 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Eps15 in U‑118‑MG Human Cell Line by Flow Cytometry.

U-118-MG human glioblastoma/astrocytoma cell line was stained with Rabbit Anti-Human Eps15 Monoclonal Antibody (Catalog # MAB8480, filled histogram) or isotype control antibody (Catalog # AB-105-C, open histogram), followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.

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Applications for Human Eps15 Antibody

Application

Recommended Usage

Immunocytochemistry

1-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Intracellular Staining by Flow Cytometry

0.25 µg/10⁶ cells
Sample: U‑118‑MG human glioblastoma/astrocytoma cell line was fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012)

Simple Western

1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line, MCF‑7 human breast cancer cell line, A431 human epithelial carcinoma cell line, and Jurkat human acute T cell leukemia cell line

Western Blot

0.1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and A431 human epithelial carcinoma cell line

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS For liquid material, refer to CoA for concentration.

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.

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: Eps15

Eps15 (Epidermal growth factor receptor substrate 15) is a 138-140 kDa member of the Eps family of proteins. Eps15 has a tripartite structure comprising an amino terminal portion, which contains three evolutionary conserved EH protein-protein interaction domains, a central putative coiled-coil region required for constitutive oligmerization, and a carboxy terminal domain containing multiple copies of the amino acid triplet aspartate-prolinephenylalanine that constitute the AP2 binding domain. The carboxy terminal domain also contains two ubiquitin interaction motifs (UIMs), the last of which is indespensible for Eps15 binding to ubiquitin. Eps15 binds to AP-2 as well as other proteins involved in endocytosis and/or synaptic vesicle recycling, such as synaptojanin1 and epsin. Furthermore, Eps15 colocalizes with markers of the plasma membrane clathrin-coated pits and vesicles.The EPS15 gene yields two isoforms that are believed to reside in distinct subcellular locations and thus implicated in different facets of endosomal trafficking. Human EPS15 has been mapped to chromosome 1p31- p32, a region displaying several non-random chromosomal abnormalities, including deletions in neuroblastoma and translocation in acute lymphoblastic and myeloid leukemias. Over aa 492-579, human EPS15 shares 76% aa identity with mouse EPS15.

Long Name

EGFR Pathway Substrate 15

Alternate Names

AF1P, MLLT5

Entrez Gene IDs

2060 (Human); 13858 (Mouse); 313474 (Rat)

Gene Symbol

EPS15

UniProt

P42566

Additional Eps15 Products

Product Documents for Human Eps15 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human Eps15 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human Eps15 Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB8480

Key Product Details

Species Reactivity

Validated:

Applications

Label

Antibody Source

View all Eps15 Antibodies »

Product Summary for Human Eps15 Antibody

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human Eps15 Antibody

Detection of Human Eps15 by Western Blot.