Human/Mouse/Rat EEA1 Biotinylated Antibody

Catalog #: BAF8047 Datasheet / COA / SDS

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BAF8047

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Human/Mouse/Rat EEA1 Biotinylated Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

EEA1 antibodies are ideal for immunocytochemistry colocalization studies in endosomes. The unconjugated antibody detects human, mouse, and rat EEA1 in Western blots.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human EEA1
Asn1249-Gln1356
Accession # Q15075

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.

Label

Biotin

Applications

Recommended Concentration

Sample

Immunocytochemistry

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

Immunocytochemistry

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EEA1 in HeLa Human Cell Line. EEA1 was detected in formaldehyde fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human/Mouse/Rat EEA1 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF8047) at 15 µg/mL overnight at 4 °C. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (orange; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to endosomes. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

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.

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: EEA1

EEA1 (Early Endosome Autoantigen 1; also known as Endosome-associated protein p162 and Zn-finger FYVE domain-containing protein 2) is a 170-180 kDa protein that serves as an identifying marker for early endosomes. It is ubiquitously expressed, and found in both the cytosol and on cellular membranes. Its activity has been described as that of a tethering factor which links endosomes to endocytic vesicles, allowing for their fusion via a SNARE complex. Normally, EEA1 exists as a homodimer in the cytoplasm and appears to make transient contacts with endosome membrane phosphatidylinositol. When endosome fusion is not required, EEA1 serves as a substrate for p97, promoting EEA1 dissociation and endosome independence. When endosome fusion is required, EEA1 interacts with NSF, resulting in its removal from a large endosome-associated complex and subsequent endosomal vesicle fusion. Human EEA1 is synthesized as a 1411 amino acid (aa) protein that contains one C2H2-type Zn finger region (aa 41-64) and one FYVE Zn finger domain (aa 1352-1410). There is one isoform variant that contains a nine aa substitution for aa 925-1411. Over aa 1249-1356, human EEA1 shares 99% aa sequence identity with mouse EEA1.

Long Name

Early Endosome Antigen 1

Entrez Gene IDs

8411 (Human); 216238 (Mouse); 314764 (Rat)

Alternate Names

early endosome antigen 1; early endosome antigen 1, 162kD; early endosome-associated protein; EEA1; Endosome-associated protein p162; MST105; MSTP105; ZFYVE2; ZFYVE2MST105; Zinc finger FYVE domain-containing protein 2