Anti-phospho-TSC2-Ser939 Antibody, clone 1G19 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1657 - recombinant, expressed in HEK 293 cells
Synonym: Tuberous sclerosis 2 protein
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Prostate (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in subsets of tubules & glomeruli of Human Prostate tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Prostate (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in subsets of tubules & glomeruli of Human Prostate tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Prostate (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in subsets of tubules & glomeruli of Human Prostate tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Kidney (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in subsets of tubules & glomeruli of Human Kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Kidney (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in subsets of tubules & glomeruli of Human Kidney tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Kidney (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in subsets of tubules & glomeruli of Human Kidney tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HEK 293 cells (A and B) and HEK 293 treated with Anisomycin (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 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 Membrane and cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HEK 293 cells (A and B) and HEK 293 treated with Anisomycin (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 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 Membrane and cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HEK 293 cells (A and B) and HEK 293 treated with Anisomycin (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 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 Membrane and cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HEK 293 cells (A and B) and HEK 293 treated with Anisomycin (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 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 Membrane and cytoplasm.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HEK 293 cells (A and B) and HEK 293 treated with Anisomycin (C and D) was performed using a 1:100 dilution of Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 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 Membrane and cytoplasm.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Constructs containing non-phospho-TSC2/Tuberin-Ser939 (Lane 1) and phospho-TSC2/Tuberin-Ser939 (Lane 2) recombinant proteins were probed with Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-TSC2-Ser939 (~48 kDa). Constructs courtesy of Dr. Jesse Rinehart, Yale University, School of Medicine.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from untreated HeLa cells (Lane 1) and HeLa cells treated with Anisomycin (25 μg/mL; 30 min) (Lane 2) were probed with Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-TSC2-Ser939 (~220 kDa).
Peptide Inhibition Assay Enhanced Validation - Recombinant Antibody Technology Lysates from HeLa cells treated with Anisomycin (25 μg/mL; 30 min) were probed with Cat. No. ZRB1657, Anti-phospho-TSC2-Ser939, clone 1G19 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-TSC2-Ser939 (~220 kDa). Lanes 1: Incubated without any peptide. 2: Incubated with phosphorylated Tuberin-TSC2-Ser939 peptide. 3: Incubated with non-phosphorylated Tuberin-TSC2-Ser939 peptide.
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 |
| clone | 1G19, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | Internal |
| 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 200.61 kDa |
| observed mol wt ~220 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_000539  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | mouse, rat |
| storage temp. | 2-8°C |
| technique(s) | immunocytochemistry: suitable |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| peptide synthesis: suitable | |
| western blot: suitable | |
| UniProt accession no. | P49815 |
| Application: | Quality Control Testing Evaluated by Western Blotting in HeLa treated with Anisomycin. Western Blotting Analysis: A 1:10,000 dilution of this antibody detected phospho-Tuberin/TSC2-Ser9 Tested applications Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected phospho-TSC2-Ser939 in Hek293 treated with Anisomycin and phospho-TSC2/Tuberin-Ser9 Immunohistochemistry (Paraffin) Analysis: A 1:1,000 dilution from a representative lot detected phospho-TSC2-Ser939 in human kidney and human prostate tissue sections. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected phospho-TSC2-Ser939 in HEK 293 treated with Anisomycin. Peptide Inhibition Assay: Target band detection in lysate from HeLa cells treated with Anisomycin was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| 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. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 10 amino acids surrounding phosphoserine 939 from the internal region of human Tuberin/TSC2. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. 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 per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 1G19 is a ZooMAb® Rabbit recombinant monoclonal antibody that detects Tuberin/TSC2. It targets an epitope within 10 amino acids surrounding phosphoserine 939. |
| 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: | Tuberin (UniProt: P49815; also known as Tuberous sclerosis 2 protein) is encoded by the TSC2 (also known as TSC4) gene (Gene ID: 7249) in human. TSC2 is a peripheral membrane protein that serves as a tumor suppressor and is expressed in multiple tissues. In complex with TSC1, it inhibits the nutrient-mediated or growth factor-stimulated phosphorylation of S6K1 and EIF4EBP1 by negatively regulating mTORC1 signaling. It acts as a GTPase-activating protein (GAP) for the small GTPase Rheb, a direct activator of the protein kinase activity of mTORC1. In its C-terminal region is a highly conserved Rap-Gap domain (aa 1531-1758) that is essential for its function. TSC2 has been reported to increase the intrinsic GTPase activity of both Rap1A and Rab5 in vitro. It can undergo phosphorylation at serine 1387, 1418 or 1420. However, these phosphorylations do not affect its interaction with TSC1. It can also undergo phosphorylation at serine 939 and threonine 1462 by Akt1 induced by growth factor stimulation. When phosphorylated and activated by AMPK under conditions of low ATP:AMP ratio, it leads to negative regulation of the mTORC1 complex. AMPK-mediated phosphorylation leads to TSC1-TSC2 complex stabilization, increasing GAP activity toward Rheb and allows turning off mTORC1 signaling. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Li, Y., et al. (2004). Trends Biochem. Sci. 29(1); 32-38; Tee, AR., et al. (2002). Proc. Natl. Acad. Sci. USA. 99(21); 13571-13576 |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |