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Product Info Summary
| SKU: | A03757-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-ATG9A Antibody Picoband®
SKU/Catalog Number
A03757-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ATG9A Antibody catalog # A03757-2. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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-ATG9A Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03757-2)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human ATG9A recombinant protein (Position: M1-D812).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A03757-2 is reactive to ATG9A in Human, Mouse
Observed Molecular Weight
94 kDa
Calculated molecular weight
94.4 kDa
Background of ATG9A
Autophagy-related protein 9A is a protein that in humans is encoded by the ATG9A gene. ATG9A is the only transmembrane ATG protein essential for autophagy. It plays a key role in the organization of the preautophagosomal structure/phagophore assembly site (PAS). It has been reported that ATG9A expression is increased in oral squamous cell carcinoma and breast cancers. The inhibition of ATG9A can lead to an inhibition of cancer cell proliferation and invasion.
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
A03757-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, 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.
Western blot, 0.25-0.5μg/ml, Human, Mouse
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human HepG2 whole cell, human A549 whole cell, human A375 whole cell, human Mcf-7 whole cell, human K562 whole cell, mouse testis tissue
ICC/IF: A549 cell
FCM: U87 cell
Validation Images & Assay Conditions
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Western blot analysis of ATG9A using anti-ATG9A antibody (A03757-2).
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 HepG2 whole cell lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: human A375 whole cell lysates,
Lane 4: human Mcf-7 whole cell lysates,
Lane 5: human K562 whole cell lysates,
Lane 6: mouse testis tissue 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-ATG9A antigen affinity purified polyclonal antibody (Catalog # A03757-2) 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 1:5000 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 ATG9A at approximately 94KD. The expected band size for ATG9A is at 94KD.
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IF analysis of ATG9A using anti-ATG9A antibody (A03757-2).
ATG9A was detected in immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-ATG9A Antibody (A03757-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of U87 cells using anti-ATG9A antibody (A03757-2).
Overlay histogram showing U87 cells stained with A03757-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ATG9A Antibody (A03757-2, 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-ATG9A Antibody Picoband® (A03757-2)
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