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Human/Mouse/Rat ATG3 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF5450

R&D Systems, part of Bio-Techne
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AF5450
AF5450-SP

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human

Applications

Validated:

Immunocytochemistry, Western Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG

Product Specifications

Immunogen

E. coli-derived recombinant human ATG3
Met1-His287
Accession # Q9NT62

Specificity

Detects endogenous human, mouse and rat ATG3 in Western blots.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat ATG3 Antibody

Detection of Human/Mouse/Rat ATG3 antibody by Western Blot.

Detection of Human/Mouse/Rat ATG3 by Western Blot.

Western blot shows lysates of human brain tissue, mouse brain tissue, and PC-12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat ATG3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5450) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for ATG3 at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
ATG3 antibody in RAW 264 by Immunocytochemistry (ICC).7 Mouse Cell Line by Immunocytochemistry (ICC).

ATG3 in RAW 264.7 Mouse Cell Line.

ATG3 was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line, untreated (lower panel) or treated with LPS (upper panel), using Sheep Anti-Human/Mouse/Rat ATG3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5450) at 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 autophagosomes. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Applications for Human/Mouse/Rat ATG3 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed RAW264.7 mouse monocyte/macrophage cell line

Western Blot

1 µg/mL
Sample: Human brain tissue, mouse brain tissue, and PC-12 rat adrenal pheochromocytoma cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. 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: ATG3

ATG3 (autophagy-related protein 3; also APG3-like and PC3-96) is a ubiquitous 45 kDa member of the ATG3 family of proteins. It functions as an E2-like enzyme during the initial stages of autophagosome formation by catalyzing the transfer of ATG7-bound ATG8 (known as LC3, GATE16 and GABA-RAP in mammals) to phosphatidylethanolamine, critical for autophagy. Human ATG3 is 314 amino acids in length and contains an active site at Cys264 that forms a thiol ester bond with the C-terminal Gly of ATG8. There are multiple potential isoform variants. Three show a 22, 28 and 35 aa substitution for aa 289‑314, respectively, while a fourth shows an alternate start site at Met88 that may be accompanied by one of the afore mentioned substitutions. Over aa 1‑287, human ATG3 shares 97.9% aa identity with mouse ATG3.

Long Name

Autophagy Related 3 Homolog

Alternate Names

APG3L, Apg3p, PC3-96

Entrez Gene IDs

64422 (Human); 67841 (Mouse); 171415 (Rat)

Gene Symbol

ATG3

UniProt

Additional ATG3 Products

Product Documents for Human/Mouse/Rat ATG3 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 ATG3 Antibody

For research use only

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