ATG12 Recombinant Rabbit Monoclonal Antibody [JE30-61]
cat.: HA721504
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
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
Images
HA721504_1.jpg 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.
HA721504_2.jpg 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.
HA721504_3.jpg 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.