Luciferase Antibody

Immunocytochemistry/Immunofluorescence: Luciferase Antibody [NB100-1677] - Human tumor xenograft expressing firefly luciferase intracranially injected into a mouse brain. Image provided via product review by verified customer.

Western Blot: Luciferase Antibody [NB100-1677] - Lane 1: Luciferase, 50ng.Luciferase antibody at 1:1000 overnight at 4C. Secondary antibody: Peroxidase goat secondary antibody at 1:40,000 for 30 min at RT. Block: incubated with blocking buffer for 30 min at RT. Predicted/Observed size: 60 kDa for Luciferase. Other band(s): None.

Immunocytochemistry/Immunofluorescence: Luciferase Antibody [NB100-1677] - In vitro expression of Luciferase by 247Nluc+ recombinant strain of Alcelaphine herpesvirus 1 (AIHV-1). Epifluoresence expression of Luciferase by AIHV-1 infected syncytia. MDBK cells were infected with the WT (i to iv) or 247Nluc+ (v to viii) strain. Five days p.i. the the syncytia were fixed with 4% paraformaldehyde in PBS, permeabilized in 0.1% NP-40 in PBS and revealed by indirect IF co-staining. See image for additional details. Image provided via product review by verified customer.

Immunohistochemistry-Paraffin: Luciferase Antibody [NB100-1677] - Analysis of human breast tumor xenograft expressing firefly luciferase subcutaneously injected into a mouse using Luciferase antibody. Primary antibody dilution 1:200. Image from verified customer review.

Immunohistochemistry-Paraffin: Luciferase Antibody [NB100-1677] - NIH3T3 cells. Cells (25,000/well) transfected with pLuc plasmid were stained with 20ug/ml NB100-1677 and 1:200 dilution of donkey anti-goat IgG-FITC (green). Cells were mounted with DAPI (blue) and visualized at 200X magnification.

Immunohistochemistry-Paraffin: Luciferase Antibody [NB100-1677] - Luciferase immunostaining was analyzed in tibias from ovx ERE-luciferase mice taken 24 h after a 17-E2 injection. Positive luciferase staining was identified in hypertrophic chondrocytes (hc) (A), lining cells (lc) (B), osteoblasts (ob) (C), a subpopulation (10%) of osteocytes (ot) (upper arrow points at a positively stained osteocyte, whereas the lower depicts a negative osteocyte) (D), and megakaryocytes (E). Faint staining was found on the periosteal surface (p) (F). No background staining was seen when omitting the primary antibody (data not shown). The bar in the lower left corner represents 25 um. pc, Proliferative chondrocyte.

Immunohistochemistry-Paraffin: Luciferase Antibody [NB100-1677] - Luciferase expression in mouse brain tissue bearing human glioma cells expressing fire fly luciferase. Image from verified customer review.

• Reactivity: Hu, Pp
• Applications: WB, ELISA, ICC/IF, IHC, IHC-Fr, IHC-P
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated

Luciferase Antibody Summary

• Immunogen: Luciferase [Photinus pyralis (Firefly)]
• Specificity: No reactivity is observed against Sea pansy (Renilla reniformis) luciferase.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Purity: Multi-step

Applications/Dilutions

• Dilutions:
  • ELISA 1:200-1:1000
  • Immunocytochemistry/Immunofluorescence 20 ug/ml
  • Immunohistochemistry 1:1000-1:5000
  • Immunohistochemistry-Frozen 1:1000 - 1:5000
  • Immunohistochemistry-Paraffin 1:1000 - 1:5000
  • Western Blot 1:1000-1:5000
• Application Notes: This antibody has been tested in Western Blot, IHC, and ELISA. Expect a band ~60kDa in appropriate cell lysates. Although not tested, this antibody would be useful in immunofluorescence, immunoprecipitation, immunocytochemistry, and most immunological methods requiring high titers and specificity.

Reactivity Notes

No reactivity is observed against Sea pansy (Renilla reniformis) luciferase.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Preservative: 0.01% Sodium Azide
• Purity: Multi-step

Notes

This product is an IgG fraction antibody purified from monospecific antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Goat Serum as well as purified and partially purified Luciferase [Photinus pyralis (Firefly)]

Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Alternate Names for Luciferase Antibody

• ec 1.13.12.7
• firefly luciferase
• Fluc
• LuC
• Luciferase
• luciferin 4 monooxygenase
• Luciferin 4-monooxygenase

Background

Luciferase is a generic term for a group of oxidative enzymes used in bioluminescence. Firefly (Photinus pyralis) and bacterial luciferase enzymes are commonly used in assay systems such as cell viability assays, reporter gene assays, and for in vivo imaging. Bacterial luciferases are flavoenzymes composed of two subunits each encoded by the luxA and luxB genes, while the firefly luciferase is a single polypeptide specified by the luc gene (1). Firefly luciferase (theoretical molecular weight: 61 kDa) oxidizes the substrate luciferin to oxyluciferin in a bioluminescent reaction requiring Mg2+ and ATP (2,3). This reaction produces a flash of yellow-green light with an emission peak around 560nm that can be detected by a luminometer (3). Firefly luciferase has become one of the more widely used reporter proteins and is an excellent tool for the study of gene expression, given that the amount of light emitted is directly proportional to luciferase activity (4).

The luciferase assay is fast and sensitive, differentiating itself from the CAT (chloramphenicol acetyltransferase) assay because it does not require a radioactive substrate.

References

1. Eun, H. (1996). Marker/Reporter enzymes. Enzymology Primer for Recombinant DNA Technology, 567-645. doi:10.1016/b978-012243740-3/50011-9

2. McNabb, D. S., Reed, R., & Marciniak, R. A. (2005). Dual luciferase assay system for rapid assessment of gene expression in Saccharomyces cerevisiae. Eukaryotic Cell, 4(9), 1539-1549. doi:10.1128/ec.4.9.1539-1549.2005

3. Fraga, H. (2008). Firefly luminescence: A historical perspective and recent developments. Photochemical & Photobiological Sciences, 7(2), 146-158. doi:10.1039/b719181b

4. Younes, A., Lukyanenko, Y. O., Lyashkov, A. E., Lakatta, E. G., & Sollott, S. J. (2011). A bioluminescence method for direct measurement of phosphodiesterase activity. Analytical Biochemistry, 417(1), 36-40. doi:10.1016/j.ab.2011.05.036

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Luciferase Antibody (NB100-1677)(23)

Publications using NB100-1677

• Trieu KG, Tsai SY, Eberl M et al. Basal cell carcinomas acquire secondary mutations to overcome dormancy and progress from microscopic to macroscopic disease Cell reports 2022-05-03 [PMID: 35508126] (ICC/IF)
• Lu X, Fong KW, Gritsina G et al. HOXB13 suppresses de novo lipogenesis through HDAC3-mediated epigenetic reprogramming in prostate cancer Nature genetics [PMID: 35468964] (MiAr)
• Noffsinger B, Witter A, Sheybani N Et al. Technical choices significantly alter the adaptive immune response against immunocompetent murine gliomas in a model-dependent manner Journal of neuro-oncology 2021-08-25 [PMID: 34432197]
• Gutierrez S, Eisenach Jc, Boada Md Seeding of breast cancer cell line (MDA-MB-231LUC+) to the mandible induces overexpression of substance P and CGRP throughout the trigeminal ganglion and widespread peripheral sensory neuropathy throughout all three of its divisions Molecular pain 2021-07-07 [PMID: 34229504] (IHC-Fr)
• Lazaro-Ibanez E, Faruqu FN, Saleh AF et al. Selection of Fluorescent, Bioluminescent, and Radioactive Tracers to Accurately Reflect Extracellular Vesicle Biodistribution in Vivo ACS Nano 2021-01-20 [PMID: 33470092] (WB)
• Kang MH, van Lieshout LP, Xu L et al. A lung tropic AAV vector improves survival in a mouse model of surfactant B deficiency Nat Commun 2020-08-09 [PMID: 32764559] (ICC/IF)
• Garg S, Wu C et al. Express ELISA for detection of mortalin. Biotechniques 2019-01-10 [PMID: 31502469] (ICC/IF)
• Chen TH, Arra M, Mbalaviele G, Swarnkar G, Abu-Amer Y. Inflammatory Responses Reprogram T(REGS) Through Impairment of Neuropilin-1 Sci Rep 2019-07-20 [PMID: 31320680] (IHC-P)
• Ohta Y, Nomura E, Shang J, et al. Enhanced oxidative stress and the treatment by edaravone in mice model of amyotrophic lateral sclerosis J Neurosci Res 2018-12-20 [PMID: 30565312] (ICC/IF)
• Singh RD, Hillestad ML, Livia C, et al. M(3)RNA Drives Targeted Gene Delivery in Acute Myocardial Infarction Tissue Eng Part A 2018-07-27 [PMID: 30047313] (ICC/IF)
• Henrich MT, Geibl FF, Lee B et al. A53T-a-synuclein overexpression in murine locus coeruleus induces Parkinson's disease-like pathology in neurons and glia Acta Neuropathol Commun 2018-05-10 [PMID: 29747690] (ICC/IF, Firefly)
• Yun Y, Oh J, Kim Y et al. Characterization of neural stem cells modified with hypoxia/neuron-specific VEGF expression system for spinal cord injury Gene Ther. 2017-11-20 [PMID: 29155421]
• Luengo-Gil G, Calvo MI, Martin-Villar E et al. Antithrombin controls tumor migration, invasion and angiogenesis by inhibition of enteropeptidase. Sci Rep 2016-06-09 [PMID: 27270881] (IHC-Fr)
• Vadrot N, Duband-Goulet I, Cabet E, et al. The p.R482W substitution in A-type lamins deregulates SREBP1 activity in Dunnigan-type familial partial lipodystrophy Hum Mol Genet 2014-12-20 [PMID: 25524705] (ICC/IF)
• Cho H, Herzka T, Zheng W, et al. RapidCaP, a novel GEM model for metastatic prostate cancer analysis and therapy, reveals myc as a driver of Pten-mutant metastasis Cancer Discov 2014-01-22 [PMID: 24444712] (IHC-P)
• Sayed D, He M, Yang Z et al. Transcriptional Regulation Patterns Revealed by High Resolution Chromatin Immunoprecipitation during Cardiac Hypertrophy. J Biol Chem 2013-01-25 [PMID: 23229551] (WB, Firefly)
• Ozdemir M, Attar A, Kuzu I. Regenerative treatment in spinal cord injury Curr Stem Cell Res Ther 2012-09-01 [PMID: 22849699]
• Yang X, Balakrishnan I, Torok-Storb B, et al. Marrow Stromal Cell Infusion Rescues Hematopoiesis in Lethally Irradiated Mice despite Rapid Clearance after Infusion Adv Hematol 2012-03-09 [PMID: 22400029] (IHC)
• Kawabata A, Baoum A, Ohta N, et al. Intratracheal administration of a nanoparticle-based therapy with the angiotensin II type 2 receptor gene attenuates lung cancer growth Cancer Res 2012-03-06 [PMID: 22389453] (IHC)
• Dewals B, Myster F, Palmeira L et al. Ex vivo bioluminescence detection of alcelaphine herpesvirus 1 infection during malignant catarrhal fever J Virol 2011-07-01 [PMID: 21593175] (ICC/IF)
• Yang G, Abate A, George AG, et al. Maturational differences in lung NF-kappaB activation and their role in tolerance to hyperoxia J Clin Invest 2004-09-03 [PMID: 15343385] (IHC)
• Timmermann L The Locus coeruleus in Parkinson's disease-from basic research to new translational perspectives Thesis (IHC)
• Ozdemir M, Attar A, Kuzu I, et al. Stem cell therapy in spinal cord injury: in vivo and postmortem tracking of bone marrow mononuclear or mesenchymal stem cells Stem Cell Rev Rep 2012-05-04 [PMID: 22552878] (ICC/IF, IHC-Fr)

FAQs for Luciferase Antibody (NB100-1677). (Showing 1 - 2 of 2 FAQs).

1. One of our customers would like additional information about your goat anti-luciferase (catalog #NB100-1677). They want to use this antibody for immunohistochemistry of paraffin-embedded sections, and would like to know if you recommend an antigen retrieval step? If so, which one is best?
   • For IHC-P with NB100-1677, we would recommend 10 mm citrate buffer (pH 6.0) based heat induced antigen retrieval at 95-99 C.
2. Hello, can I use this antibody (NB100-1677) for immunohistochemistry on rat frozen brain sections?
   • This antibody will work in IHC on frozen and paraffin embedded tissues, provided that they express Firefly Luciferase from Photinus pyralis.

Bioinformatics

• Uniprot:
  • P08659(Firefly)