Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
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Species reactivity: | Human |
Applications: | WB, IF-Cell, FC |
Clonality: | Monoclonal |
Clone number: | JE30-61 |
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: 15 kDa |
Isotype: | IgG |
Immunogen: | Synthetic peptide within Human ATG12 aa 1-50 / 140. |
Positive control: | HeLa cell lysate, HCT 116 cell lysate, HEK-293 cell lysate, SH-SY5Y cell lysate, HCT 116. |
Subcellular location: | Cytoplasm, Preautophagosomal structure membrane. |
Recommended Dilutions:
WB IF-Cell FC |
1:1,000 1:250 1:500-1:1,000 |
Uniprot #: | SwissProt: O94817 Human |
Alternative names: | Apg12 APG12-like APG12L ATG12 ATG12 autophagy related 12 homolog (S. cerevisiae) ATG12 autophagy related 12 homolog ATG12_HUMAN Autophagy 12 Autophagy-related protein 12 FBR93 HAPG12 Ubiquitin-like protein ATG12 |
Fig1:
Western blot analysis of ATG12 on different lysates with Rabbit anti-ATG12 antibody (HA721504) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HCT 116 cell lysate Lane 3: HEK-293 cell lysate Lane 4: SH-SY5Y cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 15 kDa Observed band size: 55/20 kDa Exposure time: 3 minutes; 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 (HA721504) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of HCT 116 cells labeling ATG12 with Rabbit anti-ATG12 antibody (HA721504) at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ATG12 antibody (HA721504) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. 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. |
Fig3:
Flow cytometric analysis of HCT 116 cells labeling ATG12. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721504, 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). |