ATG5 Antibody (603813)

R&D Systems | Catalog # MAB5294

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Western Blot, Simple Western

Cited:

Immunohistochemistry, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 603813
View all ATG5 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human ATG5
Asn99-Thr193
Accession # Q9H1Y0

Specificity

Detects human, mouse, and rat ATG5 in Western blots.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for ATG5 Antibody (603813)

Detection of Human, Mouse, and Rat ATG5 antibody by Western Blot.

Detection of Human, Mouse, and Rat ATG5 by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, CH-1 mouse B cell lymphoma cell line, and PC-12 rat adrenal pheochromocytoma cell line. PVDF Membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat ATG5 Monoclonal Antibody (Catalog # MAB5294) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for ATG5 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Detection of Human and Rat ATG5 antibody by Simple WesternTM.

Detection of Human and Rat ATG5 by Simple WesternTM.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and PC‑12 rat adrenal pheochromocytoma cell line, loaded at 0.2 mg/mL. A specific band was detected for ATG5 at approximately 51 kDa (as indicated) using 25 µg/mL of Mouse Anti-Human/Mouse/Rat ATG5 Monoclonal Antibody (Catalog # MAB5294). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of ATG5 by Flow Cytometry

Detection of ATG5 by Flow Cytometry

Role of Atg-5 in OXGR1 expression. CD14+ monocytes were differentiated to moMDSCs with GM-CSF/IL-6 either in the presence of alpha KG or without it (ctrl) for 4 days and then treated with LPS for the next 16-18 h. A) Epifluorescence images of cells collected 0.5h, 2h or 24h after the initial cultures and stained with anti-OXGR1 Ab, and anti-IgG:Alexa 488 and DAPI. B) Representative flow cytometry analysis of surface OXGR1 expression in control (grey histograms) and alpha KG-treated (red histograms) cells collected as in (A). C) Representative ROS measurements by DHE staining in moMDSCs pretreated with 1 µg/mL siRNAatg5 or control scrambled siRNA (encapsulated in LNPs with 0.5 µg/mL ApoE, Supplement Figure S7) for 24 h, and then differentiated with GM-CSF and IL-6, either with or without 30 mM alpha KG for 5 days and LPS for the last 16-18h. D) Representative analysis of OXGR1 expression in LPS-treated moMDSCs after the cultures as in (C). A-D) Representative data is shown from one donor, out of three different donors with similar results. E) Concentrations of IL-1 beta and IL-23 were measured in supernatants of cultures prepared as in (C). Data is shown as mean ± SEM (n=3). *p<0.05 as indicated (RM-ANOVA, Dunnett's post-test). Image collected and cropped by CiteAb from the following open publication (https://www.ijbs.com/v22p5119.htm), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of ATG5 by Western Blot

Detection of ATG5 by Western Blot

Role of autophagy in response of moMDSCs to alpha KG. A) Relative Atg5, lc3, p62, bcln and ambre mRNA expression, measured by qPCR from moMDSCs treated or not with alpha KG (30 mM) for 48h is shown from one experiment carried out in triplicates, out of 2 experiments with similar results. B) Representative wester blots of ATG5, LC3B-I, LC3B-II with GAPDH as loading control from moMDSCs treated on not with alpha KG (5mM or 30 mM) for 48h, and C) the summarized data normalized to control moMDSCs (1) are shown. D) MoMDSCs pretreated with 1 µg/mL siRNAoxgr1 or control scrambled siRNA (encapsulated in LNPs with 0.5 µg/mL ApoE, Supplement Figure S7) for 24 h, and then differentiated with GM-CSF and IL-6 with or without 30 mM alpha KG for 4 days were analysed for LC3 flux (ratio between bafilomycin-treated and non-treated cells) after staining for membrane-bond LC3 expression and the analysis on Cell Muse Analyser (see Supplementary Figure S8A for representative histograms). E) Representative western blot of LC3B-I and LC3B-II with GAPDH as loading control in moMDSCs pretreated with 1 µg/mL siRNAatg5 or control siRNA, as in D (see Supplementary Figure S8 for the equivalent LC3 flux experiment), and F) the summarized data are shown. G) IL-10 concentrations in culture supernatants of moMDSCs, pretreated with siRNAs (as in D and E), and then treated or not with alpha KG (30mM) for 5 days and LPS (100 ng/ml) for the last 16-18h, as indicated. (C,D, F,G) Data is shown as mean ± SEM of 3 independent experiments. *p < 0.05 as indicated (RM-ANOVA, Dunnett's post-test). Image collected and cropped by CiteAb from the following open publication (https://www.ijbs.com/v22p5119.htm), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for ATG5 Antibody (603813)

Application
Recommended Usage

Immunohistochemistry

8-25 µg/mL
Sample: Immersion fixed paraffin-embedded human small intestine

Simple Western

25 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and PC‑12 rat adrenal pheochromocytoma cell line

Western Blot

0.5 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line, CH‑1 mouse B cell lymphoma cell line, and PC‑12 rat adrenal pheochromocytoma cell line

Reviewed Applications

Read 1 review rated 5 using MAB5294 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: ATG5

ATG5 (Autophagy-related Protein), also known as APG5L and Apoptosis-specific Protein, is a ubiquitous 32 kDa member of the ATG family of proteins. ATG5 exists as a covalent heterodimer with ATG12 through the creation of a Lys-Gly linkage. The ATG5:ATG12 heterodimer associates noncovalently with an ATG16 multimer to generate autophagosomes. Human ATG5 is 275 amino acids in length and contains N- and C-terminal ubiquitin-like domains (aa 15‑105 and 187‑275) separated by a helix-rich linker region that contains a dimerizing Lys at position 130. There are two potential alternate start sites at Met80 and Met173. Over aa 99‑193, human ATG5 is 97% aa identical to mouse ATG5.

Long Name

ATG5 Autophagy Related 5 Homolog

Alternate Names

APG5, ASP

Entrez Gene IDs

9474 (Human); 11793 (Mouse)

Gene Symbol

ATG5

UniProt

Q9H1Y0 (Human)

Additional ATG5 Products

Product Documents for ATG5 Antibody (603813)

Certificate of Analysis

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Product Specific Notices for ATG5 Antibody (603813)

For research use only

Related Research Areas

Autophagy

Citations for ATG5 Antibody (603813)

Customer Reviews for ATG5 Antibody (603813) (1)

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  • Human/Mouse/Rat ATG5 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: CH-1 mouse B cell lymphoma cell line
    Species: Mouse
    Verified Customer | Posted 07/25/2022
    ATG5 Antibody (603813) MAB5294

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