IHC
Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissues using ATG4D antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded Human prostate cancer tissues using ATG4D antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.ICC/IF
Immunofluorescence analysis of A549 cells using ATG4D antibody (green). Blue: DAPI fluorescent DNA dye.Cells are fixed in 4% paraformaldehyde at room temperature for 20 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Rabbit IgG(H&L) (Green,OM643486), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160).IHC
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissues using ATG4D antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.IHC
Immunohistochemical analysis of paraffin-embedded Human lung cancer tissues using ATG4D antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.WB
Western blot analysis using ATG4D antibody against Mouse brain(1),Mouse intestine(2),Mouse testes(3) tissue. 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000-1:2000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S.ELISA
Coat the plate with immunizing antigen.Blue line： Immune serum; Orange line: Pre-immune serum.| Product Name | Anti-ATG4D antibody,pbs only |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Polypeptide |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ELISAICC/IFIHCWB |
| IHC:1:200-1:2000 ICC/IF:1:100-1:500 WB:1:1000-1:2000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | ATG4D |
|---|---|
| Gene Synonyms | 9830134P12Rik APG4-D APG4D Atg4dl AUTL4 HsAPG4D SPGF101 |
| Gene Full Name | autophagy related 4D cysteine peptidase |
| Gene Summary | Autophagy is the process by which endogenous proteins and damaged organelles are destroyed intracellularly. Autophagy is postulated to be essential for cell homeostasis and cell remodeling during differentiation, metamorphosis, non-apoptotic cell death, and aging. Reduced levels of autophagy have been described in some malignant tumors, and a role for autophagy in controlling the unregulated cell growth linked to cancer has been proposed. This gene belongs to the autophagy-related protein 4 (Atg4) family of C54 endopeptidases. Members of this family encode proteins that play a role in the biogenesis of autophagosomes, which sequester the cytosol and organelles for degradation by lysosomes. Alternative splicing results in multiple transcript variants. |
| Alternative Names | APG4 (ATG4) autophagy-related homolog D APG4 autophagy 4 homolog D ATG4 autophagy related 4 homolog D AUT-like 4 cysteine endopeptidase AUT-like 4 cysteine endopeptidase autophagin-4 autophagy-related 4D autophagy-related cysteine endopeptidase 4 autophagy-related protein 4 homolog D cysteine protease involved in autophagy cysteine protease involved in autophagy APG4-D |
| Molecular Weight(MW) | 53KDA |
| Cellular Localization | Cytoplasm, Mitochondrion |
| SwissProt ID | Q86TL0 |
|---|
IHC
Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissues using ATG4D antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded Human prostate cancer tissues using ATG4D antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
ICC/IF
Immunofluorescence analysis of A549 cells using ATG4D antibody (green). Blue: DAPI fluorescent DNA dye.Cells are fixed in 4% paraformaldehyde at room temperature for 20 minutes. Then, they are permeabilized with a PBS (OM750003) solution containing 0.1% Triton X-100(OM750021) at room temperature for 15 minutes. Subsequently, the cells are blocked with 10% non - immune goat serum(OM760028) at room temperature for 1 hour.The cells are incubated overnight at 4°C with the primary antibody diluted 1:100 in PBS. The secondary antibody, Omnimabs® 488 Goat Ant-Rabbit IgG(H&L) (Green,OM643486), is diluted at a ratio of 1:400 and incubated with the cells for 1 hour.Nuclear DNA is labeled with DAPI (Blue,OM643160).
IHC
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissues using ATG4D antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
IHC
Immunohistochemical analysis of paraffin-embedded Human lung cancer tissues using ATG4D antibody with DAB staining.Pre-treat the sections with heat-mediated antigen retrieval using sodium citrate buffer (pH 6.0) (OM750020) for 2 minutes. Wash the sections with ddH₂O and PBS (OM750003). Block the tissue with 10% non-immune goat serum(OM760028) at room temperature for 30 minutes. Incubate the tissue with the primary antibody diluted at a ratio of 1:1500 at 4°C overnight. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:200 and incubate for one hour. Use DAB(OM760029)as the chromogenic agent. Counterstain the tissue with hematoxylin, and mount the tissue sections with neutral gum.
WB
Western blot analysis using ATG4D antibody against Mouse brain(1),Mouse intestine(2),Mouse testes(3) tissue. 12% SDS-PAGE gel.Sample loading: 20μg /lane. Transfer the proteins onto a PVDF membrane (OM790003), and block it with TBST (OM750016) plus skimmed milk powder(OM750084) for one hour. Dilute the primary antibody with the antibody diluent (OM750012) at a ratio of 1:1000-1:2000, and incubate it overnight at 4°C. Wash the membrane three times with TBST (OM750016), 5 minutes each time. At room temperature, dilute the secondary antibody, Goat Anti-Rabbit IgG(H&L)-HRP (OM643487), at a ratio of 1:20000 and incubate for one hour. Wash the membrane three times with TBST (OM750016) again, 5 minutes each time. Use ECL (OM625701) for luminescence.staining time: 60S.
ELISA
Coat the plate with immunizing antigen.Blue line： Immune serum; Orange line: Pre-immune serum.| Application Notes | IHC:1:200-1:2000 ICC/IF:1:100-1:500 WB:1:1000-1:2000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. |
| Storage Buffer | PBS ONLY |
