ATG5 Antibody

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ProSci
Product Code: PSI-5031
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-5031-0.02mg0.02mg£150.00
Quantity:
PSI-5031-0.1mg0.1mg£449.00
Quantity:
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Overview

Host Type: Chicken
Antibody Isotype: IgY
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

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<strong>Figure 1 Western Blot Validation of ATG5 in Mouse Spleen </strong><br> Loading: 15 μg of lysates per lane. Antibodies: ATG5, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-chicken IgG HRP conjugate at 1:10000 dilution. Lane A: 1 μg/mL Lane B: 2 μg/mL
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<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: ATG5 5031 (2 μg/mL), ATG5 64-229 (2 μg/mL), actin (1 μg/mL), and GAPDH (0.02 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-chicken or goat anti-mouse IgG HRP conjugate at 1:10000 and 1:5000 dilution, respectively.
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<strong>Figure 3 Western Blot Validation of ATG5 in Human Cell Lines </strong><br> Loading: 15 μg of lysates per lane. Antibodies: ATG 5031 (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-chicken IgG HRP conjugate at 1:10000 dilution.
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<strong>Figure 4 Immunohistochemistry Validation of ATG5 in Human Spleen </strong><br> Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-ATG5 antibody (5031) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-chicken IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.
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<strong>Figure 1 Western Blot Validation of ATG5 in Mouse Spleen </strong><br> Loading: 15 μg of lysates per lane. Antibodies: ATG5, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-chicken IgG HRP conjugate at 1:10000 dilution. Lane A: 1 μg/mL Lane B: 2 μg/mL
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br> Loading: 15 μg of lysates per lane. Antibodies: ATG5 5031 (2 μg/mL), ATG5 64-229 (2 μg/mL), actin (1 μg/mL), and GAPDH (0.02 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-chicken or goat anti-mouse IgG HRP conjugate at 1:10000 and 1:5000 dilution, respectively.
<strong>Figure 3 Western Blot Validation of ATG5 in Human Cell Lines </strong><br> Loading: 15 μg of lysates per lane. Antibodies: ATG 5031 (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-chicken IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunohistochemistry Validation of ATG5 in Human Spleen </strong><br> Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-ATG5 antibody (5031) at 2.5 μg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4˚C. A goat anti-chicken IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Documents

Further Information

Additional Names:
ATG5 Antibody: ASP, APG5, APG5L, hAPG5, APG5-LIKE
Application Note:
WB: 1-2 μg/mL; IHC-P: 2.5 μg/mL.

Antibody validated: Western Blot in human and mouse samples, Immunohistochemistry in human samples. All other applications and species not yet tested.
Background:
ATG5 Antibody: Autophagy, the process of bulk degradation of cellular proteins through an autophagosomic-lysosomal pathway is important for normal growth control and may be defective in tumor cells. It is involved in the preservation of cellular nutrients under starvation conditions as well as the normal turnover of cytosolic components. This process is negatively regulated by TOR (Target of rapamycin) through phosphorylation of autophagy protein APG1. ATG5, another member of the autophagy protein family, forms a conjugate with ATG12; this conjugate has a ubiquitin-protein ligase (E3)-like activity for protein lipidation in autophagy. This conjugate also associates with innate immune response proteins such as RIG-I and VISA (also known as IPS-1), inhibiting type I interferon production and permitting viral replication in host cells.
Background References:
  • Gozuacik and Kimchi. Oncogene 2004; 23:2891-906.
  • Kisen et al. Carcinogenesis 1993; 14:2501-5.
  • Kamada et al. J. Cell. Biol. 2000; 150:1507-13.
  • Hanada et al. J. Biol. Chem.2007; 282:37298-302.
Buffer:
ATG5 Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
DISCLAIMER:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.
Homology:
Predicted species reactivity based on immunogen sequence: Rat (100%), Bovine: (94%), Pig: (94%)
Immunogen:
Anti-ATG5 antibody (5031) was raised against a 16 amino acid synthetic peptide from near the amino terminus of human ATG5.

The immunogen is located within the first 50 amino acids of ATG5.
ISOFORMS:
Human ATG5 has 2 isoforms, including isoform L (275aa, 32.4kD) and isoform S (197aa, 22.9kD. This antibody detects only human isoform L. Mouse ATG5 has 1 isoform (275aa, 32.4kD). Rat ATG5 has only one isoform identified so far (275aa, 32.4kD).
NCBI Gene ID #:
9474
NCBI Official Name:
ATG5 autophagy related 5 homolog (S. cerevisiae)
NCBI Official Symbol:
ATG5
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 32kD

Observed: 55kD (ATG5-ATG12 conjugation)
Protein Accession #:
EAW48415
Protein GI Number:
119568800
Purification:
ATG5 Antibody is affinity chromatography purified via peptide column.
Research Area:
Autophagy
SPECIFICITY:
Three isoforms of ATG5 are known to exist; this ATG5 antibody will only detect the longest isoform.
Swissprot #:
A9UGY9
User NOte:
Optimal dilutions for each application to be determined by the researcher.
VALIDATION:

Independent Antibody Validation in Cell lines (Figure 2) shows similar ATG5 expression profile in human and mouse cell lines detected by two independent anti-ATG5 antibodies that recognize different epitopes, 5031 against N-terminus domain and 64-229 against human recombinant protein.  ATG5 proteins are detected in all the tested cell lines at different expression levels by the two independent antibodies.  

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