ATG4A Recombinant Rabbit Monoclonal Antibody [JE32-31]
cat.: HA721010
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IHC-P, FC, IF-Cell |
| Clonality: | Monoclonal |
| Clone number: | JE32-31 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. 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: 45 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within human ATG4A aa 1-50/398. |
| Positive control: | K562 cell lysate, Jurkat cell lysate, mouse kidney tissue lysate, rat kidney tissue lysate, human testis tissue, human striated muscle tissue, mouse kidney tissue, PC-3M, Hela. |
| Subcellular location: | Cytoplasm. |
| Recommended Dilutions:
WB IHC-P FC IF-Cell |
1:500 1:100-1:400 1:500-1:1,000 1:200 |
| Uniprot #: | SwissProt: Q8WYN0 Human | Q8C9S8 Mouse | B1H274 Rat |
| Alternative names: | AI627006 APG4 autophagy 4 homolog A Apg4a ATG4 autophagy related 4 homolog A (S. cerevisiae) ATG4 autophagy related 4 homolog A ATG4A ATG4A_HUMAN Atg4al AUT like 2 cysteine endopeptidase AUT-like 2 cysteine endopeptidase Autl2 Autophagin 2 Autophagin-2 Autophagy related 4A cysteine peptidase Autophagy related cysteine endopeptidase 2 Autophagy related protein 4 homolog A Autophagy-related cysteine endopeptidase 2 Autophagy-related protein 4 homolog A AV169859 Cysteine protease ATG4A hAPG4A MGC107179 |
Images
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Fig1:
Western blot analysis of ATG4A on different lysates with Rabbit anti-ATG4A antibody (HA721010) at 1/500 dilution. Lane 1: K562 cell lysate, 10 µg/Lane Lane 2: Jurkat cell lysate, 10 µg/Lane Lane 3: Mouse kidney tissue lysate, 20 µg/Lane Lane 4: Rat kidney tissue lysate, 20 µg/Lane Predicted band size: 45 kDa Observed band size: 41 kDa Exposure time: 2 minutes; 10% 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 (HA721010) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-ATG4A antibody (HA721010) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721010) at 1/100 dilution for 1 hour 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. |
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Fig3:
Immunohistochemical analysis of paraffin-embedded human striated muscle tissue with Rabbit anti-ATG4A antibody (HA721010) at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721010) at 1/400 dilution for 1 hour 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. |
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Fig4:
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-ATG4A antibody (HA721010) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721010) at 1/100 dilution for 1 hour 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. |
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Fig5:
Flow cytometric analysis of PC-3M cells labeling ATG4A. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721010, 1ug/ml) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig6:
Immunocytochemistry analysis of Hela cells labeling ATG4A with Rabbit anti-ATG4A antibody (HA721010) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-ATG4A antibody (HA721010) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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