Human Eps15 Antibody

Catalog #: AF8480 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF8480
AF8480-SP

Detection of Human Eps15 by Western Blot.

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

Human Eps15 Antibody Summary

Species Reactivity

Human

Specificity

Detects human Eps15 in direct ELISAs and Western blots.

Source

Polyclonal Rabbit IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Eps15
Met492-Thr579
Accession # P42566

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

0.05 µg/mL

See below

Simple Western

2.5 µg/mL

Jurkat human acute T cell leukemia cell line and A431 human epithelial carcinoma cell line

Immunocytochemistry

5-15 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.

Scientific Data

Western Blot

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Detection of Human Eps15 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line and A431 human epithelial carcinoma cell line. PVDF membrane was probed with 0.05 µg/mL of Rabbit Anti-Human Eps15 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8480) 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.

Immunocytochemistry

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Eps15 in HeLa Human Cell Line. Eps15 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Rabbit Anti-Human Eps15 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8480) at 0.3 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 granules in the cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunocytochemistry

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Eps15 in NIH‑3T3 Mouse Cell Line. Eps15 was detected in immersion fixed NIH-3T3 mouse embryonic fibroblast cell line using Rabbit Anti-Human Eps15 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8480) at 10 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.

Simple Western

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Detection of Human Eps15 by Simple Western^TM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line and A431 human epithelial carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Eps15 at approximately 141 kDa (as indicated) using 2.5 µg/mL of Rabbit Anti-Human Eps15 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8480). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 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: Eps15

EPS15 (Epidermal growth factor receptor substrate 15) is a 138-140 kDa member of the EPS15 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

Entrez Gene IDs

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

Alternate Names

AF1P; AF-1P; ALL1 fused gene from chromosome 1; epidermal growth factor receptor pathway substrate 15; epidermal growth factor receptor substrate 15; Eps15; MLLT5; Protein AF-1p; Protein Eps15