ATG12 Recombinant Rabbit Monoclonal Antibody [JE30-61]
Catalog# HA721504
ATG12 Recombinant Rabbit Monoclonal Antibody [JE30-61]
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WB
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IF-Cell
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FC
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Human
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unconjugated
Overview
Product Name
ATG12 Recombinant Rabbit Monoclonal Antibody [JE30-61]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human ATG12 aa 1-50 / 140.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, FC
Target Molecular Weight
Predicted band size: 15 kDa
Positive Control
HeLa cell lysate, HCT 116 cell lysate, HEK-293 cell lysate, SH-SY5Y cell lysate, HCT 116.
Conjugation
unconjugated
Clone Number
JE30-61
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000
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IF-Cell
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1:250
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FC
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1:500-1:1,000
Target
Function
Autophagy related 12 is a protein that in humans is encoded by the ATG12 gene. 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). Autophagy requires the covalent attachment of the protein Atg12 to ATG5 through a ubiquitin-like conjugation system. The Atg12-Atg5 conjugate then promotes the conjugation of ATG8 to the lipid phosphatidylethanolamine. Atg12 was found to be involved in apoptosis. This protein promotes apoptosis through an interaction with anti-apoptotic members of the Bcl-2 family.
Background References
1. Lin TY et al. BIRC5/Survivin is a novel ATG12-ATG5 conjugate interactor and an autophagy-induced DNA damage suppressor in human cancer and mouse embryonic fibroblast cells. Autophagy. 2020 Jul
2. Keulers TG et al. ATG12 deficiency results in intracellular glutamine depletion, abrogation of tumor hypoxia and a favorable prognosis in cancer. Autophagy. 2022 Aug
Subcellular Location
Cytoplasm, Preautophagosomal structure membrane.
UNIPROT
Synonyms
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
ExpandImages
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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. -
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. -
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).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"