ATG5 Antibody (YA599)

ATG5 Antibody (YA599) is a Rabbit-derived and non-conjugated IgG monoclonal antibody, targeting to ATG5.

ATG5, a key participant in autophagic vesicle formation, undergoes conjugation with ATG12 through a ubiquitin-like system involving ATG7 as an E1-like activating enzyme and ATG10 as an E2-like conjugating enzyme. The resulting ATG12-ATG5 conjugate serves as an E3-like enzyme essential for lipidation of ATG8 family proteins and their association with vesicle membranes. Beyond its role in autophagy, ATG5 contributes to mitochondrial quality control post-oxidative damage, impacting cellular longevity. It plays a critical role in lymphocyte development, vital for the survival and proliferation of both B and T lymphocytes, and is necessary for optimal antigen processing and presentation for MHC II. Additionally, ATG5 is involved in maintaining axon morphology, normal adipocyte differentiation, and promoting primary ciliogenesis through autophagic pathways. Moreover, it may play a crucial role in apoptotic processes, occurring downstream of caspase activity, and participates in IFN-gamma-induced autophagic cell death through interaction with FADD.

Free

Q9H1Y0

Predicted band size: 32 kDa; Observed band size: 55 kDa

Protein A affinity purified.

Cytoplasm, Membrane.

Non-conjugated

Unmodified

AB_3102265

Cell Biology

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Rat, Mouse, Monkey

WB: 1:1,000-1:5,000; ICC/IF: 1:50-1:200; IHC-P: 1:50-1:100; IP: 1-2μg/sample;FC: 1:1,000;IF-Tissur:1:50-1:200

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  Western blot analysis of extracts from NIH3T3(lane 2(20μg) , C2C12 (lane 3(20μg), Hela (lane 4(20μg) and HEK293(lane 5(20μg) using ATG5 (HY-P80022) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of NIH/3T3 cells labeling ATG5 with ATG5 Antibody (HY-P80022)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with ATG5 Antibody (HY-P80022) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of NIH/3T3 cells labeling ATG5 with ATG5 Antibody (HY-P80022) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with ATG5 Antibody (HY-P80022) at 1/100 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).

WB, IHC-P, ICC/IF, IP, FC, IF-Tissue

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human ATG5.AA range:1-50.