ATG5 Recombinant Rabbit Monoclonal Antibody [SN73-07]
cat.: ET1611-38
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Rat, Mouse, Monkey
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP
Clonality: Monoclonal
Clone number: SN73-07
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 32 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human ATG5 aa 1-50 / 275.
Positive control: 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.
Subcellular location: Cytoplasm, Membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  IP

1:1,000-1:5,000
1:50-1:200
1:50-1:200
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: Q9H1Y0 Human | Q99J83 Mouse | Q3MQ06 Rat
Alternative names: APG 5 APG 5L APG5 APG5 autophagy 5 like APG5 like APG5-like APG5L Apoptosis specific protein Apoptosis-specific protein ASP ATG 5 Atg5 ATG5 autophagy related 5 homolog ATG5_HUMAN Autophagy protein 5 Autophagy related 5 hAPG5 Homolog of S Cerevisiae autophagy 5 OTTHUMP00000040507
Images
ET1611-38_1.jpg Fig1: Western blot analysis of ATG5 on different lysates with Rabbit anti-ATG5 antibody (ET1611-38) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: NIH/3T3 cell lysate (15 µg/Lane)
Lane 2: C2C12 cell lysate (15 µg/Lane)
Lane 3: Neuro-2a cell lysate (15 µg/Lane)
Lane 4: PC-12 cell lysate (15 µg/Lane)
Lane 5: Mouse brain tissue lysate (15 µg/Lane)
Lane 6: Rat brain tissue lysate (15 µg/Lane)

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

Exposure time: 35 seconds;

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 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1611-38_2.jpg Fig2: 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 Fig3: 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 Fig4: 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 Fig5: 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 Fig6: 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 Fig7: 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 Fig8: 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
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.