Anti-ADAR1 Antibody, clone 1I4-H1 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1814 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (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. ZRB1814, Anti-ADAR1, clone 1I4-H1 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear/cytoplasmic staining was observed in neurons, glial cells & endothelium of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (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. ZRB1814, Anti-ADAR1, clone 1I4-H1 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear/cytoplasmic staining was observed in neurons, glial cells & endothelium of human cerebral cortex tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (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. ZRB1814, Anti-ADAR1, clone 1I4-H1 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Nuclear/cytoplasmic staining was observed in neurons, glial cells & endothelium of human cerebral cortex tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:100 dilution of Cat. No. ZRB1814, Anti-ADAR1, clone 1I4-H1 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 nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:100 dilution of Cat. No. ZRB1814, Anti-ADAR1, clone 1I4-H1 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 nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells was performed using a 1:100 dilution of Cat. No. ZRB1814, Anti-ADAR1, clone 1I4-H1 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 nucleus.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from NTERA-2 (Lane 1), HeLa (Lane 2), and L6 (Lane 3) cells were probed with Cat. No. ZRB1814, Anti-ADAR1, clone 1I4-H1 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 ADAR1 (~150 and 110 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 10 mg/mL of non-specific GST protein (Run 1) or ADAR1 recombinant protein fragment (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1814, Anti-ADAR1, clone 1I4-H1 ZooMAb® Rabbit Monoclonal. The binding of this antibody to ADAR1 recombinant protein displayed a KD of 3.4 x 10-8 whereas binding to non-specific GST protein displayed a KD of 2.1 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 | 1I4-H1, 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 136.01 kDa |
| observed mol wt ~150 and 110 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_001102  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human, 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. | P55265 |
| Application: | Quality Control Testing Evaluated by Western Blotting in HeLa cell lysate. Western Blotting Analysis: A 1:1,000 dilution of this antibody detected ADAR1 in HeLa cell lysate. Tested applications Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected ADAR1 in NTERA-2 and L6 cell lysates. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected ADAR1 in human cerebral cortex tissue sections. Affinity Binding Assay: A representative lot of this antibody bound ADAR1 with a KD of 3.4 x 10-8 in an affinity binding assay. Immunocytochemistry Analysis: A 1:100 dilution from a representative lo detected ADAR1 in HeLa cells. 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: | His-tagged recombinant fragment corresponding to 243 amino acids from the internal region of human Double-stranded RNA-specific adenosine deaminase (ADAR1). |
| 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 114-H1 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects Double-stranded RNA-specific adenosine deaminase (ADAR1). It targets an epitope within the internal 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: | Double-stranded RNA-specific adenosine deaminase (UniProt: P55265; also known as EC:3.5.4.37, DRADA, 136 kDa double-stranded RNA-binding protein, p136, Interferon-inducible protein 4, IFI-4, K88DSRBP, ADAR1) is encoded by the ADAR (also known as ADAR1, DSRAD, G1P1, IFI4) gene (Gene ID: 103) in human. In mammals three different ADAR proteins have been described: ADAR1, -2, and -3. ADAR1 is a ubiquitously expressed homodimeric enzyme that catalyzes the hydrolytic deamination of adenosine to inosine (A to I) in double-stranded RNA (dsRNA). A-to-I editing is shown to predominantly occur in noncoding, repetitive elements and short interspersed nuclear elements (SINEs). This A-to-I editing affects gene expression and function in several ways, including mRNA translation by changing codons and hence the amino acid sequence of proteins, RNA stability by changing sequences during viral RNA replication, and RNA structure-dependent activities such as microRNA production or targeting or protein-RNA interactions. Homodimerization of ADAR1 is shown to be essential for its catalytic activity. Five isoforms of ADAR1 have been described that are produced by alternative promoter usage and alternative splicing. Isoform 5, which can form heterodimers with ADAR2, is expressed at higher levels in astrocytomas and its expression increases with severity of the tumor. Loss of ADAR1 is shown to overcome resistance to PD-1 checkpoint blockade caused by inactivation of antigen presentation by tumor cells and sensitizes tumors to interferons. This ZooMAbZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Ishizuka, JJ., et al. (2019). Nature. 565(7737); 43-48; Cho, DSC., et al. (2003). J. Biol. Chem. 278(19); 17093-17102; George, CX., and Samuel, CE (1999). Proc. Natl. Acad. Sci. USA. 96(8); 4621-4626). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |