Anti-ATG9A Antibody, clone 5B11, ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1648 - recombinant, expressed in HEK 293 cells
Synonym: APG9-like 1; Autophagy-related protein 9A
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) negative control (A) and human spleen (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:10 dilution of Cat. No. ZRB1648, Anti-ATG9A, clone 5B11 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/membranous staining was observed in white pulp and to a lesser extent, red pulp of human spleen tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human salivary gland (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in glandular cells of human salivary gland tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) negative control (A) and human spleen (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:10 dilution of Cat. No. ZRB1648, Anti-ATG9A, clone 5B11 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/membranous staining was observed in white pulp and to a lesser extent, red pulp of human spleen tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human salivary gland (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in glandular cells of human salivary gland tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) negative control (A) and human spleen (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:10 dilution of Cat. No. ZRB1648, Anti-ATG9A, clone 5B11 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/membranous staining was observed in white pulp and to a lesser extent, red pulp of human spleen tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) negative control (A) and human kidney (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:10 dilution of Cat. No. ZRB1648, Anti-ATG9A, clone 5B11 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/membranous staining was observed at varying intensities in human kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) negative control (A) and human kidney (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:10 dilution of Cat. No. ZRB1648, Anti-ATG9A, clone 5B11 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/membranous staining was observed at varying intensities in human kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human salivary gland (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in glandular cells of human salivary gland tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) negative control (A) and human kidney (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:10 dilution of Cat. No. ZRB1648, Anti-ATG9A, clone 5B11 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit and DAB. Cytoplasmic/membranous staining was observed at varying intensities in human kidney tissue sections.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1648, Anti-ATG9A, clone 5B11 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1648, Anti-ATG9A, clone 5B11 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of HepG2 cell line was performed using a 1:1,000 dilution of Cat. No. ZRB1648, Anti-ATG9A, clone 5B11 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:100 dilution of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:100 dilution of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of A431 cells was performed using a 1:100 dilution of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:100 dilution of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:100 dilution of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:100 dilution of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cells was performed using a 1:100 dilution of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cells was performed using a 1:100 dilution of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytosol.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of NIH3T3 cells was performed using a 1:100 dilution of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the cytosol.
Western Blotting Enhanced Validation-Recombinant Antibody Technology Lysates from MCF7 cells (A) and HepG2 cells (B) were probed with Cat. No. ZRB1648, Anti-ATG9A, clone 5B11 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates ATG9A (~110 kDa).
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from C6 (Lane 1), NIH3T3 (Lane 2), A431 (Lane 3), and HeLa (Lane 4) cells were probed with Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates PHGDH (~56 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or PHGDH peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal. The binding of this antibody to PHGDH peptide displayed a KD of 3.8 x 10-7, whereas binding to non-specific control peptide displayed a KD of 1.0 x 10-3.
Flow Cytometry Enhanced Validation - Recombinant Antibody Technology Staining of one million C6 cells was performed using 1 μg of Cat. No. ZRB1647, Anti-PHGDH, clone 1E6 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (Grey histogram), followed by a PE-conjugated Donkey Anti-Rabbit IgG secondary antibody.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit (recombinant) |
| clone | 5B11, monoclonal |
| recombinant monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 94.45 kDa |
| observed mol wt ~110 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | canine, bovine |
| storage temp. | 2-8°C |
| technique(s) | immunocytochemistry: suitable using 1:100 |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): 1:10-1:100 | |
| western blot: suitable using 1:1,000 | |
| UniProt accession no. | Q7Z3C6  |
| Application: | Anti-ATG9A, clone 5B11 ZooMAb, Cat. No. ZRB1648, is a recombinant Rabbit Monoclonal Antibody that specifically targets Autophagy-related protein 9A and has been tested for use in Immunocytochemistry, Immunohistochemistry (Paraffin), and Western Blotting. |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected ATG9A in HepG2 cell lysate. Immunohistochemistry (Paraffin) Analysis: A 1:10 dilution from a representative lot detected ATG9A in human spleen and human kidney tissue sections. Immunocytochemistry Analysis: A 1:1000 dilution from a representative lot detected ATG9A in HepG2 cells. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 16 amino acids from the C-terminal region of human Autophagy-related protein 9A. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS with 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 5B11 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects human Autophagy-related protein 9A. It targets an epitope with in 16 amino acids from the C-terminal region. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Autophagy-related protein 9A (UniProt: Q7Z3C6; also known as APG9-like 1, ATG9) is encoded by the ATG9A (also known as APG9L1) gene (Gene ID: 79065) in human. ATG9A is a multi-pass membrane protein that is involved in autophagy and cytoplasm to vacuole transport vesicle formation. It plays a key role in the organization of the preautophagosomal structure/phagophore assembly site. Among core autophagy-related proteins, ATG9A is the only multi-spanning transmembrane protein. Both its N- and C-terminal domains are exposed to the cytoplasm. Its N-terminal region (aa 1-36) is important in its role in autophagosome formation and this region is required for both autophagy and its Golgi localization. This region also contains sorting motifs known as the di-acidic motif (aa 5-7), a tyrosine-based motif (aa 8-11), and a di-leucine-based motif (aa 22-27). In mammalian cells, ATG9A is mainly localized to the trans-Golgi network and the endosomal system. It is shown to cycle between them though vesicle trafficking. ATG9A travels through the trans-Golgi network and the endosomal system under nutrient-rich conditions, however, upon induction of autophagy it transiently localizes to the autophagosome. Under amino acid starvation or upon rapamycin treatment, it is shown to redistribute from a juxtanuclear clustered pool to a dispersed peripheral cytosolic pool. Mutant forms of ATG9A that lack both the tyrosine-based and di-leucine-based sorting motifs exhibit reduced Golgi localization and abnormal accumulation on the recycling endosomes under conditions of nutrient deprivation. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Imai, K., et al. (2016). J. Cell Sci. 129(20); 3781-3791). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |