WB
Western blot analysis of ATG12 on different lysates with Rabbit anti-ATG12 antibody 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of HCT 116 cells labeling ATG12 with Rabbit anti-ATG12 antibody 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 at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of HCT 116 cells labeling ATG12. Cells were fixed and permeabilized. Then stained with the primary antibody ( 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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | ATG12 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human ATG12 aa 1-50 / 140. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFWB |
| WB:1:1000 ICC:1:250 FC:1:500-1:1000 |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | ATG12 |
|---|---|
| Gene Synonyms | APG12 FBR93 APG12L HAPG12 |
| Gene Full Name | autophagy related 12 |
| Gene Summary | Autophagy is a process of bulk protein degradation in which cytoplasmic components, including organelles, are enclosed in double-membrane structures called autophagosomes and delivered to lysosomes or vacuoles for degradation. ATG12 is the human homolog of a yeast protein involved in autophagy (Mizushima et al., 1998 [PubMed 9852036]).[supplied by OMIM, Mar 2008] |
| Alternative Names | Apg12 antibody APG12-like antibody APG12L antibody ATG12 antibody ATG12 autophagy related 12 homolog (S. cerevisiae) antibody ATG12 autophagy related 12 homolog antibody ATG12_HUMAN antibody Autophagy 12 antibody Autophagy-related protein 12 antibody FBR93 antibody Apg12 antibody APG12-like antibody APG12L antibody ATG12 antibody ATG12 autophagy related 12 homolog (S. cerevisiae) antibody ATG12 autophagy related 12 homolog antibody ATG12_HUMAN antibody Autophagy 12 antibody Autophagy-related protein 12 antibody FBR93 antibody HAPG12 antibody Ubiquitin-like protein ATG12 antibody |
| Molecular Weight(MW) | 15kDa(Observed band size: 55/20kDa) |
| Cellular Localization | Cytoplasm, Preautophagosomal structure membrane. |
WB
Western blot analysis of ATG12 on different lysates with Rabbit anti-ATG12 antibody 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of HCT 116 cells labeling ATG12 with Rabbit anti-ATG12 antibody 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 at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HCT 116 cells labeling ATG12. Cells were fixed and permeabilized. Then stained with the primary antibody ( 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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:1000 ICC:1:250 FC:1:500-1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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. |
