antibody

Anti-ATG5 Antibody [SN73-07]

ET1611-38TR

20 uL

99 USD

monoclonal

domestic rabbit

nonconjugated

SN73-07

human, mouse, rat

western blot, immunoprecipitation, immunohistochemistry - paraffin section

Anti-ATG5 Antibody [SN73-07]

Human,Rat,Mouse,Monkey

WB,IF-Cell,IF-Tissue,IHC-P,IP

Non-conjugated

Synthetic peptide within Human ATG5 aa 1-50 / 275.

SwissProt: Q9H1Y0 Human;SwissProt: Q99J83 Mouse;SwissProt: Q3MQ06 Rat

Rabbit

SN73-07

IgG

20 uL

99 USD

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

Liquid

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Protein A affinity purified.

Recombinant Rabbit monoclonal Antibody

NIH/3T3 cell lysate, C2C12 cell lysate, Neuro-2a cell lysate, PC-12 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, HeLa cell lysate, A431 cell lysate, Hela, MCF-7, human lung carcinoma tissue.

Predicted band size: 32 kDa

Cytoplasm, Membrane.

1 mg/mL.

WB: 1:1,000-1:5,000 ;IF-Cell: 1:50-1:200 ;IF-Tissue: 1:50-1:200 ;IHC-P: 1:50-1:100 ;IP: Use at an assay dependent concentration.

ET1611-38_2.jpg

Western blot analysis of ATG5 on different lysates with Rabbit anti-ATG5 antibody (ET1611-38) at 1/1,000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: A431 cell lysate (20 µg/Lane) Lane 3: PC-12 cell lysate (20 µg/Lane) Lane 4: NIH/3T3 cell lysate (20 µg/Lane) Lane 5: Neuro-2a cell lysate (20 µg/Lane) Predicted band size: 32 kDa Observed band size: 55 kDa Exposure time: 1 minute; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-38) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

ET1611-38_3.jpg

All lanes: Western blot analysis of ATG5 with anti-ATG5 antibody [SN73-07] (ET1611-38) at 1/1,000 dilution. Lane 1: Wild-type Huh7 whole cell lysate. Lane 2: ATG5 knockout Huh7 whole cell lysate. ET1611-38 was shown to specifically react with ATG5 in wild-type Huh7 cells. No band was observed when ATG5 knockout sample was tested. Wild-type and ATG5 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-ATG5 antibody (ET1611-38, 1/1,000) and Anti-GAPDH antibody (ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Cell lysate was provided by Ubigene Biosciences (Ubigene Biosciences Co., Ltd., Guangzhou, China).

ET1611-38_4.jpg

All lanes: Western blot analysis of ATG5 with anti-ATG5 antibody [SN73-07] (ET1611-38) at 1/1,000 dilution. Lane 1: Wild-type Vero-E6 whole cell lysate. Lane 2: ATG5 knockout Vero-E6 whole cell lysate. ET1611-38 was shown to specifically react with ATG5 in wild-type Vero-E6 cells. No band was observed when ATG5 knockout sample was tested. Wild-type and ATG5 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-ATG5 antibody (ET1611-38, 1/1,000) and Anti-GAPDH antibody (ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Cell lysate was provided by Ubigene Biosciences (Ubigene Biosciences Co., Ltd., Guangzhou, China).

ET1611-38_5.jpg

ICC staining of ATG5 in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-38, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

ET1611-38_6.jpg

ICC staining of ATG5 in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1611-38, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

ET1611-38_7.jpg

Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-ATG5 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1611-38, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

ET1611-38_8.jpg

ATG5 was immunoprecipitated in 0.2mg HeLa cell lysate with ET1611-38 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1611-38 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: Rabbit IgG instead of ET1611-38 in HeLa cell lysate Lane 3: ET1611-38 IP in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 1 minute 21 seconds