Anti-GLS Antibody, clone 1D16 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1798 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
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. ZRB1798, Anti-GLS, clone 1D16 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Punctate cytoplasmic staining was observed in tubules 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. ZRB1798, Anti-GLS, clone 1D16 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Punctate cytoplasmic staining was observed in tubules 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. ZRB1798, Anti-GLS, clone 1D16 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Punctate cytoplasmic staining was observed in tubules of human kidney tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of SH-SY5Y cells was performed using a 1:1,000 dilution of Cat. No. ZRB1798, Anti-GLS, clone 1D16 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitochondria.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of SH-SY5Y cells was performed using a 1:1,000 dilution of Cat. No. ZRB1798, Anti-GLS, clone 1D16 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitochondria.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of SH-SY5Y cells was performed using a 1:1,000 dilution of Cat. No. ZRB1798, Anti-GLS, clone 1D16 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa FluorZooMAb® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the mitochondria.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Mouse brain (Lane 1) and Rat brain (Lane 2) tissue lysates were probed with Cat. No. ZRB1798, Anti-GLS, clone 1D16 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 GLS (~60 kDa).
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from PC12 (Lane 1), SH-SY5Y (Lane 2), and HepG2 (Lane 3) cells were probed with Cat. No. ZRB1798, Anti-GLS, clone 1D16 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 GLS (~55-60 kDa).
Western Blotting Analysis: Lysates from mouse hepatocytes, control and siRNA treated (24h and 48h), were probed with Cat. No. ZRB1798, Anti-GLS, clone 1D16 (0.4 μg/mL) and re-probed with anti- β-actin to verify loading. Knockdown of GLS1 was observed with the GLS1 siRNA treatment, but not in lysates from control cells. (Data courtesy of María Mercado-Gómez and Teresa Cardoso Delgado, Liver Disease Lab, CIC bioGUNE, Spain).
Western Blotting Analysis: Liver tissue membranes from mice fed a control diet (Ctrl) or a choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) were probed with Cat. No. ZRB1798, Anti-GLS, clone 1D16 (0.4 μg/mL) and re-probed with anti- β-actin to verify loading. Mice fed CDAHFD showed significantly higher expression of GLS1. (Data courtesy of María Mercado-Gómez and Teresa Cardoso Delgado, Liver Disease Lab, CIC bioGUNE, Spain).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-specific control peptide (Run 1) or Glutaminase (GLS) peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1798, Anti-GLS, clone 1D16 ZooMAb® Rabbit Monoclonal. The binding of this antibody to Glutaminase (GLS) peptide displayed a KD of 2.4 x 10-6 whereas binding to non-specific control peptide displayed a KD of 3.2 x 10-3.
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 | 1D16, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | N-terminal |
| 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 73.46 kDa |
| observed mol wt ~60 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_055720  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | rat, human, mouse |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | O94925 |
| Application: | Quality Control Testing Evaluated by Western Blotting in Rat brain tissue lysate. Western Blotting Analysis: A 1:10,000 dilution of this antibody detected GLS in Rat brain tissue lysate. Tested applications Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected GLS in lysates from Mouse brain tissue and from PC12, SH-SY5Y, and HepG2 cells. Affinity Binding Assay:: A representative lot of this antibody bound GLS with a KD of 2.4 x 10-6 in an affinity binding assay. Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected GLS in SH-SY5Y cells. Immunohistochemistry (Paraffin) Analysis: A 1:1,000 dilution from a representative lot detected GLS in human kidney tissue sections. 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 23 amino acids from the N-terminal region of human Glutaminase (GLS). |
| 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 1D16 is a ZooMAb® Rabbit recombinant monoclonal antibody that detects kidney mitochondrial form of human glutaminase. |
| 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: | Glutaminase kidney isoform, mitochondrial (UniProt: O94925; also known as EC:3.5.1.2, GLS, K-glutaminase, L-glutamine amidohydrolase) is encoded by the GLS (also known as GLS1, KIAA0838) gene (Gene ID: 2744) in human. GLS catalyzes the first reaction in the primary pathway for the renal catabolism of glutamine. It is primarily expressed in the brain and kidney and plays an essential role in generating energy for metabolism, synthesizing the brain neurotransmitter glutamate, and maintaining acid-base balance in the kidney. Following its synthesis, it is found in cytoplasm and in the mitochondria. The 74-kDa cytosolic precursor is reported to translocate into the mitochondria with the help of a mitochondrial transit peptide (aa 1-54) and is processed to yield the mature 68 kDa (aa 55-669) and 65-kDa (aa 73-669)subunits. During normal acid-base balance, the kidney extraction of plasma glutamine is low, however, during metabolic acidosis, about one third of the plasma glutamine is extracted and metabolized in a single pass through kidneys. Three isoforms of glutaminase have been described in humans that are produced via alternate splicing. Isoforms 1 and 3 are activated by phosphate, while the isoform 3 lacks catalytic activity. Isoform 1 is highly expressed in brain and kidney, but is not detected in liver. On the other hand, isoform 3 is highly expressed in astrocytoma, ganglioglioma, ependymoma. Lower levels of isoform 3 are reported in placenta, lung, pancreas, and kidney, but it is not detected in liver. Isoform 2 is expressed in cardiac and skeletal muscle. Mutations in GLS gene are known to cause developmental and epileptic encephalopathy 71 (DEE71)s that is characterized by refractory seizures, neurodevelopmental impairment, and poor prognosis. This ZooMAbZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Rumping, L., et al. (2019). JAMA Neurol. 76(3); 342-350; Elgadi, KM., et al. (1999). Physiol. Genomics 1(2); 51-62). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |