PA1-18285

antibody from Invitrogen Antibodies

Targeting: ATG5 APG5, APG5L, ASP, hAPG5

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-18285

Provider

Invitrogen Antibodies

Product name

ATG5 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Reconstitute with 500 µL of distilled water. After reconstitution, store aliquots at -20ºC for a higher stability, and at 4ºC with an appropriate antibacterial agent.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

500 µg

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles. Glycerol (1:1) may be added for added stability.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of APG5 was performed by loading 30 µg of HEK293T whole cell lysate per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat dry milk in TBST for 1 hour at room temperature. The membrane was probed with an APG5 polyclonal antibody (Product # PA1-18285) at a dilution of 1:200 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Detection was performed using ECL substrate. Data courtesy of the Innovators Program.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of APG5 (red) in HEK293T cells. Cells fixed with 4% formaldehyde were permeabilized and blocked with 1X PBS containing 5% BSA and 0.3% Triton X-100 for 1 hour at room temperature. Cells were probed with an APG5 polyclonal antibody (Product # PA1-18285) at a dilution of 1:20 overnight at 4°C in 1X PBS containing 1% BSA and 0.3% Triton X-100, washed with 1X PBS, and incubated with a fluorophore-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:200 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken at 20X magnification. Data courtesy of the Innovators Program.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunoprecipitation of APG5 was performed on HEK293T cells. Antigen-antibody complexes were formed by incubating 500 µg of HEK293T whole cell lysate (in 500 µL volume) with 5 µL of an APG5 Antibody (Product # PA1-18285) overnight at 4°C. The immune complexes were captured on 30 µL of protein G agarose, washed extensively, and eluted with 6X non-reducing Laemmli buffer. Samples were resolved on a 12% SDS-PAGE, transferred to a PVDF membrane, and blocked with 5% milk in TBST for 1 hour at room temperature. The membrane was probed with an APG5 polyclonal antibody (Product # PA1-18285) at a dilution of 1:200 overnight at 4øC, washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Chemiluminescent detection was performed using ECL substrate. Data courtesy of the Innovators Program.