Anti-ATG4A Antibody Picoband™

Product Name

Anti-ATG4A Antibody Picoband™

View all ATG4A Antibodies

SKU/Catalog Number

A06539-2

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-ATG4A Antibody Picoband™ catalog # A06539-2. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-ATG4A Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A06539-2)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na₂HPO₄, 0.05mg NaN₃.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence at the N-terminus of human ATG4A, which shares 75% amino acid (aa) sequence identity with both mouse and rat ATG4A.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A06539-2 is reactive to ATG4A in Human

Applications

A06539-2 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

50 kDa

Calculated molecular weight

45.378kDa

Background of ATG4A

Cysteine protease ATG4A is an enzyme that in humans is encoded by the ATG4A gene. It is mapped to Xq22.3. 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 encodes a member of the autophagin protein family. The encoded protein is also designated as a member of the C-54 family of cysteine proteases.

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.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

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.

Western blot, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of ATG4A using anti-ATG4A antibody (A06539-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human PC-3 whole cell lysates,
Lane 4: human A431 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ATG4A antigen affinity purified polyclonal antibody (Catalog # A06539-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ATG4A at approximately 50KD. The expected band size for ATG4A is at 45KD.

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Figure 2. IHC analysis of ATG4A using anti-ATG4A antibody (A06539-2).
ATG4A was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ATG4A Antibody (A06539-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of ATG4A using anti-ATG4A antibody (A06539-2).
ATG4A was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ATG4A Antibody (A06539-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of PC-3 cells using anti-ATG4A antibody (A06539-2).
Overlay histogram showing PC-3 cells stained with A06539-2 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATG4A Antibody (A06539-2,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For ATG4A (Source: Uniprot.org, NCBI)

Alternative Names

APG4 autophagy 4 homolog A (S. cerevisiae); Apg4a; ATG4 autophagy related 4 homolog A (S. cerevisiae); ATG4A; Atg4al; Autl2; AUT-like 2 cysteine endopeptidase; AUT-like 2, cysteine endopeptidase (S. cerevisiae); Autophagin 2; autophagin-2; cysteine endopeptidase; cysteine protease ATG4A; EC 3.4.22; EC 3.4.22.-; hAPG4A ATG4A APG4A, AUTL2 autophagy related 4A cysteine peptidase cysteine protease ATG4A|APG4 autophagy 4 homolog A|ATG4 autophagy related 4 homolog A|AUT-like 2, cysteine endopeptidase|autophagin 2|autophagy-related cysteine endopeptidase 2|autophagy-related protein 4 homolog A|hAPG4A

For more info on ATG4A, check out the ATG4A Infographic

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In this infographic, you will see the following information for ATG4A: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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