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Product Info Summary
| SKU: | M06539-1 |
|---|---|
| Size: | 80 µl |
| Reactive Species: | Human |
| Host: | Mouse |
| Application: | Flow Cytometry, IF, IHC-P, WB |
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Product info
Product Name
Anti-ATG4A Antibody
SKU/Catalog Number
M06539-1
Size
80 µl
Description
Boster Bio Anti-ATG4A Antibody (Catalog # M06539-1). Tested in WB, Flow Cytometry, IHC-P, IF application(s). This antibody reacts with Human.
Storage & Handling
Maintain refrigerated at 2-8°C for up to 2 weeks. For long-term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.
Cite This Product
Anti-ATG4A Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # M06539-1)
Host
Mouse
Contents
Purified monoclonal antibody supplied in PBS with 0.09% (W/V) sodium azide.
Clonality
Monoclonal
Clone Number
1458CT808.66.25.69
Isotype
IgG2b,k
Immunogen
This ATG4A antibody is generated from a mouse immunized with a recombinant protein.
Reactive Species
M06539-1 is reactive to ATG4A in Human
Calculated molecular weight
45.4 kDa
Background of ATG4A
Cysteine protease required for the cytoplasm to vacuole transport (Cvt) and autophagy. Cleaves the C-terminal amino acid of ATG8 family proteins 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. Preferred substrate is GABARAPL2 followed by MAP1LC3A and GABARAP. Has also an activity of delipidating enzyme for the PE-conjugated forms.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
M06539-1 is guaranteed for Flow Cytometry, IF, IHC-P, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
IF: 1:25
WB: 1:500
IHC-P: 1:25
FC: 1:25
Validation Images & Assay Conditions
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervical epithelial adenocarcinoma cell line) cells labeling ATG4A with M06539-1 at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-mouse IgG secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoplasm staining on HeLa cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin at 1/100 dilution (red).The nuclear counter stain is DAPI (blue).
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Western blot analysis of lysate from K562 cell line, using ATG4A Antibody. M06539-1 was diluted at 1:500. A goat anti-mouse IgG H&L(HRP) at 1:10000 dilution was used as the secondary antibody. Lysate at 20μg.
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M06539-1 staining ATG4A in human brain sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.
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Overlay histogram showing Hela cells stained with M06539-1 (green line). The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (M06539-1, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Mouse IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed at 1/400 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was mouse IgG2b (1g/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.
Specific Publications For Anti-ATG4A Antibody (M06539-1)
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