Immunofluorescence analysis of ATG4B was done on 70% confluent log phase Rapamycin treated HeLa cells (treated with 100nM of Rapamycin A for 4 hours) . The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ATG14 Rabbit ABfinity™ Antibody (710702) at 2 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing nuclear localization. Panel e shows untreated HeLa cells. Panel f shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||full-length protein corresponding to Human ATG4b|
|Contains||0.09% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5µg/million cells|
|Western Blot (WB)||1-2µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody is predicted to react with monkey, mouse, rat, guinea pig and bovine based on sequence homology.
ABfinity™ recombinant oligoclonal antibodies are rabbit antibody pools generated from isolated DNA libraries. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ oligoclonal antibodies recognize multiple epitopes and therefore exhibit greater sensitivity and because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Cysteine protease ATG4B is an enzyme which is coded by ATG4B gene. Autophagy is a critical cellular homeostatic process that controls the turnover of damaged organelles and protein. ATG4B is a deubiquitin-like protease that plays dual roles in the core machinery of autophagy. ATG4B, cleaves the C-terminal amino acid of ATG8 family proteins MAP1LC3, GABARAPL1, GABARAPL2 and GABARAP, to reveal a C-terminal glycine. Exposure of the glycine at the C-terminus is essential for ATG8 proteins conjugation to phosphatidylethanolamine (PE) and insertion to membranes, which is necessary for autophagy. ATG4B is mainly expressed in the skeletal muscle, brain, heart, liver and pancreas.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.