ATG5/APG5L Rabbit Polyclonal Antibody

• Catalog Number: orb5704
• Category: Antibodies
• Description: ATG5/APG5L Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Equine, Gallus, Porcine, Rabbit
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human APG5L (101-200/275aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 32 kDa
• Target: ATG5
• UniProt ID: Q9H1Y0
• RRID: AB_10926251
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: ATG5; APG 5; APG 5L; APG5; APG5 autophagy 5 like;
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Li, Yue et al. Autophagy contributes to 4-Amino-2-Trifluoromethyl-Phenyl Retinate-induced differentiation in human acute promyelocytic leukemia NB4 cells Toxicol Appl Pharmacol, 319, (2017)

PubMed: 28130038

Cao, Wei et al. Reducing-Autophagy Derived Mitochondrial Dysfunction during Resveratrol Promotes Fibroblast-Like Synovial Cell Apoptosis Anat Rec (Hoboken), 301, 1179-1188 (2018)

PubMed: 29461680

Blank control: A431. Primary Antibody (green line): Rabbit Anti-ATG5 antibody (orb5704), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF647, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (human gastric), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (ATG5) Polyclonal Antibody, Unconjugated (orb5704) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human liver carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (ATG5) Polyclonal Antibody, Unconjugated (orb5704) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Mouse brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (ATG5) Polyclonal Antibody, Unconjugated (orb5704) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat kidney), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (ATG5/APG5L) Polyclonal Antibody, Unconjugated (orb5704) at 1:200 overnight at 4°C, followed by a conjugated Goat Anti-rabbit IgG antibody for 90 minutes, and DAPI for nuclei staining.

Sample: Lane 1: Mouse Cerebrum tissue lysates, Lane 2: Mouse Kidney tissue lysates, Lane 3: Rat Cerebrum tissue lysates, Primary: Anti-ATG5/APG5 (orb5704) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 32 kDa, Observed band size: 47 kDa.