Anti-Linear Ubiquitin Antibody, clone 1E3 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB2114 - recombinant, expressed in HEK 293 cells
Product Type: Chemical
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells (A and B) and HeLa cells serum starved and treated overnight with 25 nM MG132 (C and D) were performed using a 1:1,000 dilution of Cat. No. ZRB2114, Anti-Linear Ubiquitin, clone 1E3 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 nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells (A and B) and HeLa cells serum starved and treated overnight with 25 nM MG132 (C and D) were performed using a 1:1,000 dilution of Cat. No. ZRB2114, Anti-Linear Ubiquitin, clone 1E3 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 nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells (A and B) and HeLa cells serum starved and treated overnight with 25 nM MG132 (C and D) were performed using a 1:1,000 dilution of Cat. No. ZRB2114, Anti-Linear Ubiquitin, clone 1E3 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 nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells (A and B) and HeLa cells serum starved and treated overnight with 25 nM MG132 (C and D) were performed using a 1:1,000 dilution of Cat. No. ZRB2114, Anti-Linear Ubiquitin, clone 1E3 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 nucleus.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of HeLa cells (A and B) and HeLa cells serum starved and treated overnight with 25 nM MG132 (C and D) were performed using a 1:1,000 dilution of Cat. No. ZRB2114, Anti-Linear Ubiquitin, clone 1E3 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 nucleus.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Recombinant Linear Ub (Lane 1), recombinant Ub K11 (Lane 2), recombinant Ub K48 (Lane 3), and recombinant Ub K63 (Lane 4) were probed with Cat. No. ZRB2114, Anti-Linear Ubiquitin, clone 1E3 ZooMAb® Rabbit Monoclonal (1:20,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system.
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 10 mg/mL of non-specific GST protein (Run 1) or recombinant Linear Ubiquitin (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB2114, Anti-Linear Ubiquitin, clone 1E3 ZooMAb® Rabbit Monoclonal. The binding of this antibody to Linear Ubiquitin, protein displayed a KD of 4.5 x 10-8 whereas binding to non-specific GST protein displayed a KD of 1.8 x 10-5.
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 | 1E3, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | Unknown |
| 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 Variable |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_061828.1  |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| immunocytochemistry: suitable | |
| western blot: suitable | |
| UniProt accession no. | P0CG47 and P0CG48 |
| Application: | Quality Control Testing Evaluated by Western Blotting with linear Ub, Ub K11, Ub K48, and Ub K63. Western Blotting Analysis: A 1:20,000 dilution of this antibody detected Ubiquitin in Linear Ub, but not Ub K11, Ub K48, Ub K63. Tested applications Affinity Binding Assay: A representative lot of this antibody bound Ubiquitin with a KD of 4.5 x 10-8 in an affinity binding assay. Immunocytochemistry Analysis: A 1:1,000 dilution from a representative lot detected Ubiquitin in HeLa treated with MG132. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Application: | Anti-Linear Ubiquitin, clone 1E3 ZooMAb®, Cat. No. ZRB2114, is a recombinant Rabbit monoclonal antibody that detects Ubiquitin and is tested for use in Affinity Binding Assay, Immunocytochemistry, and Western Blotting. |
| 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: | Recombinant protein corresponding to all linear ubiquitin linkages. |
| 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 1E3 is a ZooMAb® rabbit recombinant monoclonal antibody that specifically detects linear ubiquitins |
| 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: | Ubiquitin (Ub) is initially produced as a 229 amino acids Polyubiquitin-B (UniProt: P0CG47) precursor protein encoded by the UBB gene (Gene ID: 7314) or a 685 amino acids Polyubiquitin-C precursor protein (UniProt: P0CG48) encoded by the UBC gene (Gene ID: 7316) in human. Ub exists either covalently attached to another protein, or free (unanchored). When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Ub is linked covalently via its carboxyl terminus (Gly76) to usually Lys residues in target proteins. In a given target Lys residue can be linked to one single Ub molecule (monoubiquitylated) or to a chain of Ub molecules (polyubiquitylated). In a polyUb chain, Ub molecules can be linked through one of the seven Lys residues (K6, K11, K27, K29, K33, K48, and K63) or through the Ub N-terminus Met1 residue (which generates linear chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked. For example, lysine 6-linked may be involved in DNA repair; lysine 11-linked is involved in endoplasmic reticulum-associated degradation (ERAD) and in cell-cycle regulation, and lysine 29-linked is involved in lysosomal degradation. Lysine 48-linked chains mark proteins for proteasomal degradation, while lysine 63-linked chains are involved in endocytosis, DNA-damage responses, and in signaling leading to activation of the transcription factor NF-kappa-B. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |