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Human/Mouse/Rat EEA1 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF8047

R&D Systems, part of Bio-Techne
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AF8047
AF8047-SP

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse

Applications

Validated:

Immunocytochemistry, Western Blot

Cited:

Immunocytochemistry, Immunohistochemistry

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG

Product Specifications

Immunogen

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

Specificity

Detects human, mouse, and rat EEA1 in Western blots.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat EEA1 Antibody

Detection of Human, Mouse, and Rat EEA1 antibody by Western Blot.

Detection of Human, Mouse, and Rat EEA1 by Western Blot.

Western blot shows lysates of JAR human choriocarcinoma cell line, Hepa 1-6 mouse hepatoma cell line, and Sheep Anti-Rat-2 rat embryonic fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat EEA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8047) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for EEA1 at approximately 170 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
EEA1 antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

EEA1 in HeLa Human Cell Line.

EEA1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human/Mouse/Rat EEA1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8047) at 1.7 and 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to early endosomes. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Applications for Human/Mouse/Rat EEA1 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line

Western Blot

1 µg/mL
Sample: JAR human choriocarcinoma cell line, Hepa 1‑6 mouse hepatoma cell line, and Rat‑2 rat embryonic fibroblast cell line

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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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: EEA1

EEA1 (Early Endosome Autoantigen 1; also 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

Alternate Names

MST105, MSTP105, ZFYVE2

Entrez Gene IDs

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

Gene Symbol

EEA1

UniProt

Additional EEA1 Products

Product Documents for Human/Mouse/Rat EEA1 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human/Mouse/Rat EEA1 Antibody

For research use only

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