Anti-TIGIT Antibody, clone 1D20 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1454 - recombinant, expressed in HEK 293 cells
Synonym: T-cell immunoreceptor with Ig and ITIM domains; V-set and immunoglobulin domain-containing protein 9; V-set and transmembrane domain-containing protein
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 Dilution of Cat. No. ZRB1454-25UL, Anti-TIGIT, clone 1D20 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 Dilution of Cat. No. ZRB1454-25UL, Anti-TIGIT, clone 1D20 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 Dilution of Cat. No. ZRB1454-25UL, Anti-TIGIT, clone 1D20 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1454, Anti-TIGIT, clone 1D20 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, membrane.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1454, Anti-TIGIT, clone 1D20 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, membrane.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of Jurkat cells was performed using a 1:100 dilution of Cat. No. ZRB1454, Anti-TIGIT, clone 1D20 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, membrane.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of K562 cells was performed using a 1:100 dilution of Cat. No. ZRB1454, Anti-TIGIT, clone 1D20 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, membrane.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of K562 cells was performed using a 1:100 dilution of Cat. No. ZRB1454, Anti-TIGIT, clone 1D20 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, membrane.
Immunocytochemistry Enhanced Validation-Recombinant Antibody Technology Immunofluorescent analysis of K562 cells was performed using a 1:100 dilution of Cat. No. ZRB1454, Anti-TIGIT, clone 1D20 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, membrane.
Western Blotting Enhanced Validation-Recombinant Antibody Technology Lysates from Jurkat cells (Lane 1), Rat Spleen (Lane 2), Mouse Thymus (Lane 3), and Human Spleen (Lane 4) were probed with Cat. No. ZRB1454, Anti-TIGIT, clone 1D20 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 TIGIT (~40 kDa).
Affinity Binding Assay Enhanced Validation-Recombinant Antibody Technology 100 μg/mL of TIGIT peptide (Run 1) or a non-specific peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1454, Anti-TIGIT, clone 1D20 ZooMAb® Rabbit Monoclonal. The binding of this antibody to CXCR4 peptide displayed a KD value of 3.6 x 10^-10 whereas its binding to non-specific peptide displayed a KD of 7.5 x 106.
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 | 1D20, 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 26.32 kDa |
| observed mol wt 40 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, mouse, rat |
| 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. | Q495A1  |
| Application: | Anti-TIGIT, clone 1D20 ZooMAb, Cat. No. ZRB1454, is a recombinant Rabbit monoclonal antibody that specifically targets TIGIT and is tested for use in Affinity Binding Assay, Immunocytochemistry, Immunohistochemistry (Paraffin), and Western Blotting. |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected TIGIT in Rat Spleen, Mouse Thymus, and Human Spleen tissue lysates. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected TIGIT in Jurkat and K562 cells. Immunohistochemistry (Paraffin) Analysis: A 1:1,000 dilution from a representative lot detected TIGIT in human tonsil tissue sections. Affinity Binding Assay: A representative lot of this antibody bound TIGIT peptide with a KD of value of 3.6 x 10-9 in an affinity binding assay. 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. Learn more about ZooMAb here. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 16 amino acids from the N-terminal, extracellular domain of human T-cell immunoreceptor with Ig and ITIM domains (TIGIT). |
| 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: | 0.3 mg/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 1D20 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects human T-cell immunoreceptor with Ig and ITIM domains (TIGIT). It targets an epitope within 16 amino acids from the extracellular domain. |
| 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: | T-cell immunoreceptor with Ig and ITIM domains (UniProt: Q495A1; also known as V-set and immunoglobulin domain-containing protein 9, V-set and transmembrane domain-containing protein 3, TIGIT) is encoded by the TIGIT (also known as VSIG9, VSTM3) gene (Gene ID: 201633) in human. TIGIT is a single-pass type I membrane protein that binds with high affinity to the poliovirus receptor (PVR) forming a heterotetrameric assembly of two TIGIT and two PVR molecules. This causes increased secretion of IL10 and decreased secretion of IL12B and suppresses T-cell activation by promoting the generation of mature immunoregulatory dendritic cells. It is expressed at low levels on peripheral memory and regulatory CD4+ T-cells and NK cells and is up-regulated following activation of these cells. It is also expressed on many cell types important in cancer, and its mechanism of action may depend on the cell type studied. TIGIT also serves as a functional ligand and induces dendritic cells to acquire a tolerogenic phenotype via ligation of PVR. TIGIT -/- mice, when challenged, develop severe experimental autoimmune encephalomyelitis compared to wild-type mice. TIGIT is synthesized with a signal peptide (aa 1-21), which is subsequently cleaved off to generate the mature form that contains an extracellular domain (aa 22-141), a transmembrane domain (aa 142-162), and a cytoplasmic domain (aa 163-244). The cytoplasmic domain contains an immunoreceptor tyrosine-based inhibitor motif (ITIM; aa 229-234), which is involved in the modulation of cellular responses. The IgV domain (aa 22-124) of TIGIT possesses distinctive (V/I)(S/T)Q, AX6G, and T(F/Y)PX1G sub-motifs that mediate a ′lock and key′ trans-interaction with cis-homodimers of PVR. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Manieri, NA., et al. (2017). Trends Immunol. 38(1); 20-28). |
| WGK Germany | WGK 1 |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |