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ATG4B Antibody (YA4591)

(Synonyms: APG4B; AUTL1)
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Based on 1 Customer Validation

ATG4B Antibody (YA4591) is a Mouse-derived and non-conjugated IgG1 monoclonal antibody, targeting to ATG4B.

For research use only. We do not sell to patients.
  • Host:

    Mouse

  • Isotype:

    IgG1

  • Application:

    WB, IHC-P, FC, ELISA

  • Reactivity :

    Human

  • Formulation:

    Supplied in PBS with 0.05% sodium azide

  • Conjugation:
    Non-conjugated
  • Experimental Validation Results for ATG4B Antibody (YA4591)
  • Experimental Validation Results for ATG4B Antibody (YA4591)
  • ATG4B Antibody (YA4591) Product Image
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Tel: 609-228-6898 E-mail: [email protected] [email protected]

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Applications

Application
WB Info
WB: Western Blot
IHC-P Info
IHC-P: Immunohistochemistry-Paraffin
FC Info
FC: Flow Cytometry
ELISA Info
ELISA: Enzyme Linked Immunosorbent Assay
Dilution Ratio 1:500-1:2000 1:200-1:1000 1:200-1:400 1:10000

Product Details

Description

ATG4B Antibody (YA4591) is a Mouse-derived and non-conjugated IgG1 monoclonal antibody, targeting to ATG4B.

  • Host Mouse
  • Clonality Monoclonal
  • Species Reactivity
    Human
  • Observed Molecular Weight
    Observed band size: 44 kDa Info
    Note: Due to possible protein modifications or aggregation, the molecular weight should be confirmed by actual measurement, and the predicted value is for reference only.
  • Calculated Molecular Weight Predicted band size: 44 kDa
Immunogen

Purified recombinant fragment of human ATG4B (AA: 1-221) expressed in E. Coli.

Purification

affinity purified.

Conjugation

Non-conjugated

Modification

Unmodified

Isotype

IgG1

Product Properties

  • Appearance

    Solution

  • Formulation

    Supplied in PBS with 0.05% sodium azide

  • Concentration

    Batch-dependent, Please check the COA for the concentration of each lot. Check Lot Concentration

  • Storage & Stability

    Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

  • Shipping

    Shipping with blue ice.

Verification Images

  • Experimental Validation Results for ATG4B Antibody (YA4591)
    Western blot analysis of extracts from HepG2(lane 1(40μg) ),Hela(lane 2(40μg) ),Ramos(lane 3(40μg) ),Jurkat(lane 4(40μg) ),PC-12(lane 5(40μg)and NIH/3T3(lane 6(40μg) ) using ATG4B antibody. Proteins were transferred to a NC membrane and blocked with 5% Skim milk in TBST for 2 hour at room temperature. The primary antibody ( 1/1000) and Loading control antibody (GAPDH，1/1000) was used in 5% Skim milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for ATG4B Antibody (YA4591)
    Flow cytometric analysis of 1X10^6 Hela cells labeling ATG4B Antibody(red). Cells were fixed with 4% paraformaldehyde and permeabilised with 0.2% Triton X-100. Then stained with the primary antibody at 1/400 dilution overnight at 4℃.Alexa Fluor® 488 Goat Anti-mouse IgG H&L (invitrogen A11001) was used as the secondary antibody at 1/1,000 dilution for 45 minutes at room temperature. Mouse IgG Isotype Control (Invitrogen 14-4714-B2, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (gray).

Background

  • Function

    ATG4B is a Cysteine protease that plays a key role in autophagy by mediating both proteolytic activation and delipidation of ATG8 family proteins. Required for canonical autophagy (macroautophagy), non-canonical autophagy as well as for mitophagy. The protease activity is required for proteolytic activation of ATG8 family proteins: cleaves the C-terminal amino acid of ATG8 proteins MAP1LC3A, MAP1LC3B, MAP1LC3C, 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. Protease activity is also required to counteract formation of high-molecular weight conjugates of ATG8 proteins (ATG8ylation): acts as a deubiquitinating-like enzyme that removes ATG8 conjugated to other proteins, such as ATG3. In addition to the protease activity, also mediates delipidation of ATG8 family proteins. Catalyzes delipidation of PE-conjugated forms of ATG8 proteins during macroautophagy. Also involved in non-canonical autophagy, a parallel pathway involving conjugation of ATG8 proteins to single membranes at endolysosomal compartments, by catalyzing delipidation of ATG8 proteins conjugated to phosphatidylserine (PS). Compared to other members of the family (ATG4A, ATG4C or ATG4C), constitutes the major protein for proteolytic activation of ATG8 proteins, while it displays weaker delipidation activity than other ATG4 paralogs. Involved in phagophore growth during mitophagy independently of its protease activity and of ATG8 proteins: acts by regulating ATG9A trafficking to mitochondria and promoting phagophore-endoplasmic reticulum contacts during the lipid transfer phase of mitophagy[1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21].

  • Subcellular Localization

    Cytoplasm; Cytoplasm, cytosol; Cytoplasmic vesicle, autophagosome; Endoplasmic reticulum; Mitochondrion

  • Isoforms & Post-Translational Modification

    Q9Y4P1 has 5 isomers: Q9Y4P1-1: 44294 Da (predicted); Q9Y4P1-2: 52529 Da (predicted); Q9Y4P1-3: 31042 Da (predicted); Q9Y4P1-4: 37037 Da (predicted); Q9Y4P1-6: 42622 Da (predicted).
    Phosphorylation at Ser-383 and Ser-392 promotes autophagy by increasing protein delipidation activity without affecting proteolytic activation of ATG8 proteins (PubMed:26378241). Phosphorylation at Ser-316 by ULK1 inhibits autophagy by decreasing both proteolytic activation and delipidation activities (PubMed:28821708). Phosphorylation at Ser-316 is dephosphorylated by protein phosphatase 2A (PP2A) (PubMed:28821708). Phosphorylation at Ser-34 by AKT2 promotes its hydrolase activity, leading to increased proteolytic activation and delipidation of ATG8 family proteins (PubMed:30443548). Phosphorylation at Ser-34 by AKT1 promotes mitochondrial localization and inhibition of the F1F0-ATP synthase activity, leading to elevation of mitochondrial reactive oxygen species (ROS) (PubMed:29165041);Ubiquitinated by RNF5, leading to its degradation by the proteasome;S-nitrosylation at Cys-189 and Cys-292 in response to high glucose decreases both proteolytic activation and delipidation activities;O-glycosylated by OGT, leading to increase protease activity, thereby promoting the proteolytic activation of ATG8 family proteins;Forms reversible intrachain disulfide bonds in response to oxidative stress (PubMed:31880198). Forms interchain disulfide bonds, leading to formation of homooligomers in response to oxidation (PubMed:31880198)

  • Subunit

    Interacts with PFKP; promoting phosphorylation of ATG4B at Ser-34 (PubMed:33607258). Interacts with GBP7 (By similarity)

  • SwissProt ID

    Q9Y4P1

  • Gene ID
    23192 [NCBI]
  • Synonyms

    APG4B; AUTL1

ATG4B Antibody (YA4591) Related Classifications

MOQ
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100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedChemExpress Validation
     Western blot analysis of extracts from HepG2(lane 1(40μg) ),Hela(lane 2(40μg) ),Ramos(lane 3(40μg) ),Jurkat(lane 4(40μg) ),PC-12(lane 5(40μg)and NIH/3T3(lane 6(40μg) ) using ATG4B antibody. Proteins were transferred to a NC membrane and blocked with 5% Skim milk in TBST for 2 hour at room temperature. The primary antibody ( 1/1000) and Loading control antibody (GAPDH，1/1000) was used in 5% Skim milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.

  • 1/16

    MedChemExpress Validation
     Flow cytometric analysis of 1X10^6 Hela cells labeling ATG4B Antibody(red). Cells were fixed with 4% paraformaldehyde and permeabilised with 0.2% Triton X-100. Then stained with the primary antibody at 1/400 dilution overnight at 4℃.Alexa Fluor® 488 Goat Anti-mouse IgG H&L (invitrogen A11001) was used as the secondary antibody at 1/1,000 dilution for 45 minutes at room temperature. Mouse IgG Isotype Control (Invitrogen 14-4714-B2, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (gray).

  • ATG4B Antibody (YA4591) Product Image
    1/16
  • Experimental Validation Results for ATG4B Antibody (YA4591)
  • Experimental Validation Results for ATG4B Antibody (YA4591)
  • ATG4B Antibody (YA4591) Product Image
  • Experimental Validation Results for ATG4B Antibody (YA4591)
    1/1
    MedChemExpress Validation
     Western blot analysis of extracts from HepG2(lane 1(40μg) ),Hela(lane 2(40μg) ),Ramos(lane 3(40μg) ),Jurkat(lane 4(40μg) ),PC-12(lane 5(40μg)and NIH/3T3(lane 6(40μg) ) using ATG4B antibody. Proteins were transferred to a NC membrane and blocked with 5% Skim milk in TBST for 2 hour at room temperature. The primary antibody ( 1/1000) and Loading control antibody (GAPDH，1/1000) was used in 5% Skim milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.
  • Experimental Validation Results for ATG4B Antibody (YA4591)
    1/1
    MedChemExpress Validation
     Flow cytometric analysis of 1X10^6 Hela cells labeling ATG4B Antibody(red). Cells were fixed with 4% paraformaldehyde and permeabilised with 0.2% Triton X-100. Then stained with the primary antibody at 1/400 dilution overnight at 4℃.Alexa Fluor® 488 Goat Anti-mouse IgG H&L (invitrogen A11001) was used as the secondary antibody at 1/1,000 dilution for 45 minutes at room temperature. Mouse IgG Isotype Control (Invitrogen 14-4714-B2, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (gray).
  • Experimental Validation Results for ATG4B Antibody (YA4591)
  • Experimental Validation Results for ATG4B Antibody (YA4591)
  • ATG4B Antibody (YA4591) Product Image
    MedChemExpress Validation

    Western blot analysis of extracts from HepG2(lane 1(40μg) ),Hela(lane 2(40μg) ),Ramos(lane 3(40μg) ),Jurkat(lane 4(40μg) ),PC-12(lane 5(40μg)and NIH/3T3(lane 6(40μg) ) using ATG4B antibody. Proteins were transferred to a NC membrane and blocked with 5% Skim milk in TBST for 2 hour at room temperature. The primary antibody ( 1/1000) and Loading control antibody (GAPDH，1/1000) was used in 5% Skim milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature.

  • ATG4B Antibody (YA4591) Product Image
    MedChemExpress Validation

    Flow cytometric analysis of 1X10^6 Hela cells labeling ATG4B Antibody(red). Cells were fixed with 4% paraformaldehyde and permeabilised with 0.2% Triton X-100. Then stained with the primary antibody at 1/400 dilution overnight at 4℃.Alexa Fluor® 488 Goat Anti-mouse IgG H&L (invitrogen A11001) was used as the secondary antibody at 1/1,000 dilution for 45 minutes at room temperature. Mouse IgG Isotype Control (Invitrogen 14-4714-B2, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (gray).

  • ATG4B Antibody (YA4591) Product Image WB
  • ATG4B Antibody (YA4591) Product Image FC