Luciferase Antibody (Luci 21 1-107) - BSA Free

Novus Biologicals | Catalog # NB600-307

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Firefly

Cited:

Mouse, Firefly

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # Luci 21 1-107

Format

BSA Free
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Product Specifications

Immunogen

This Luciferase Antibody (Luci 21 1-107) was developed against luciferase protein from Photinus pyralis (North American firefly). [UniProt# P08659].

Reactivity Notes

Photinus pyralis (North American firefly).

Localization

Peroxisome

Specificity

This Luciferase Antibody (Luci 21 1-107) is specific for Luciferase, recognizing a peptide consisting of the first 258 amino acids. Further epitope mapping has not been done at this time.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Scientific Data Images for Luciferase Antibody (Luci 21 1-107) - BSA Free

Western Blot: Luciferase Antibody (Luci 21 1-107)BSA Free [NB600-307]

Western Blot: Luciferase Antibody (Luci 21 1-107)BSA Free [NB600-307]

Luciferase-Antibody-Luci-21-1-107-Western-Blot-NB600-307-img0006.jpg
Immunohistochemistry: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307]

Immunohistochemistry: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307]

Immunohistochemistry: Luciferase Antibody (Luci 21 1-107) [NB600-307] - Detection of Luciferase expression in CD34+ cells by Immunohistochemistry. Cytospin slides prepared from transduced CD34+ cells after 3 days of culture were stained with monoclonal anti-Luciferase antibody. Luciferase-positive cells have green cytoplasm; nuclei stained with DAPI are blue. Nontransduced, cultured CD34+ cells were used as a negative control. Original magnification, 40X. Wang, X. et al., Dynamic tracking of human hematopoietic stem cell. Blood. 102(10): 3478-3482, 2003.
Western Blot: Luciferase Antibody (Luci 21 1-107)BSA Free [NB600-307]

Western Blot: Luciferase Antibody (Luci 21 1-107)BSA Free [NB600-307]

Western Blot: Luciferase Antibody (Luci 21 1-107) [NB600-307] - WB analysis of NB600-307 on Luciferase Protein (NB810-74573)
Luciferase Antibody (Luci 21 1-107) - BSA Free

Western Blot: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] -

Western Blot: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] - TTFields application leads to increased autophagic flux.a Ultra-structural STEM electron microscopy analysis of U-87 MG (upper panel) & A172 (lower panel) cells treated with TTFields for 48 h. Autophagosomes (blue arrows) & autolysosomes (green arrows) are indicated. b U-87 MG & A172 cells were either left untreated or were treated with TTFields for 24–72 h. CQ (20 µM) was added 4 h before cells were collected. Samples were immunoblotted for LC3 & GAPDH. Upper panel: representative blots. Lower panel: densitometric quantification of immunoblot signal, showing an average of at least three independent experiments (0.01 < *P < 0.05, Student’s t-test). c U-87 MG & A172 cells were either left untreated or were treated with TTFields for 48 h. CQ (20 µM) was added at the last 3 h of treatment & cells were fixed & stained with anti-LC3 Ab (green) & DAPI (blue). Upper panel: representative images. Original magnifications: × 40. Lower panel: quantification of LC3 intensity, presented as average intensity per cell (**P < 0.01, Student’s t-test). d U-87 MG cells were either left untreated or were treated with TTFields (48 h) or with vinblastin 25 nM (0.5 h). CQ (20 µM) was added at the last 3 h of treatment & cells were fixed & stained with anti-LC3 (green), LAMP1 (red), & DAPI (blue) (upper panel). Arrowheads indicate the strongest colocalization staining in each cell. Intensity histograms of LAMP1 & LC3 fluorescent signal calculated from the region of interest indicated by the white bar (lower panel). Representative images are shown Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30341282), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Luciferase Antibody (Luci 21 1-107) - BSA Free

Western Blot: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] -

Western Blot: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] - Induction of autophagy by TTFields is AMPK dependent.a U-87 MG & A172 cells either left untreated or treated w/ TTFields for indicated time points. Immunoblot analysis of GFP78 protein. Numeric values represent fold increase in GRP78 signal, normalized to loading control (GAPDH), relative to untreated control. b Quantification of intracellular ATP levels in U-87 MG cells either left untreated or treated w/ TTFields for 72 h.Results presented as average ATP concentration (nmol/2 × 106 cells) from 3 independent experiments (*P < 0.01, Student’s t-test). c U-87 MG & A172 cells either left untreated or treated w/ TTFields for indicated time points. Immunoblot analysis of pAMPK & pULK1proteins. GAPDH used as loading control. (5D-F) U-87 MG cells transfected w/ AMPK-targeting siRNA (siAMPK) or w/ siRNA sham vector (siVector), & incubated for 48 h w/ or w/out TTFields application. CQ 20 µM added for last 4 h of treatment where indicated. d (left panel) Immunoblot analysis of LC3 & AMPK. Numeric values represent fold-change in LC3-II signal, normalized to GAPDH signal, relative to respective control. d (right panel) CQ-treated cells fixed & stained for LC3 (green) & DAPI (blue), original magnifications: × 40. e Cell count of siAMPK- or siVector-expressing cells after TTFields treatment. (0.01 < *P < 0.05, Student’s t-test, n = 3). f siVector- & siAMPK-transfected U-87 MG cells either left untreated or treated w/ TTFields for 48 h. Cells then fixed & stained for cleaved caspase-3 (green) & DAPI (blue) (left panel). Images from each treatment analyzed manually & fraction of cleaved caspase-3-positive cells calculated for at least 200 cells from each group (right panel) (**P < 0.01, Student’s t-test, n = 2) Image collected & cropped by CiteAb from following publication (https://pubmed.ncbi.nlm.nih.gov/30341282), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Luciferase Antibody (Luci 21 1-107) - BSA Free

Western Blot: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] -

Western Blot: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] - TTFields induce autophagy in glioma cell lines.a U-87 MG & A172 cells were either left untreated or treated with TTFields at the last 24 h, 48 h, or 72 h of culturing. All cultures were plated on the same time, incubated overnight to allow cell attachment, & collected 72 h afterwards. Cells were collected, lysed, & samples were analyzed using immunoblotting for LC3 & GAPDH. Upper panel: representative blots. Lower panel: densitometric quantification of immunoblot signal, showing an average of at least three independent experiments (0.01 < *P < 0.05, **P < 0.01, Student’s t-test). b Paraffin-embedded sections from sham- or TTFields-treated rats were stained with anti-LC3 Ab (green) & DAPI (blue). Representative images are presented. c Quantification of LC3 intensity, presented as fold increase from corresponding control (*P < 0.05, Student’s t-test) Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30341282), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Luciferase Antibody (Luci 21 1-107) - BSA Free

Immunocytochemistry/ Immunofluorescence: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] -

Immunocytochemistry/ Immunofluorescence: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] - Immunofluorescence imaging of luciferase protein as a reporter of hybrid formation. Entire primary tumor & lung sections were imaged via tile scanning, & each image of the scan was carefully analyzed to confirm or refute positive staining for luciferase. The luciferase signal was considered a positive signal if it was above background levels associated with negative controls & corresponded to the cytoplasm of a cell with a nucleus. Rare luciferase-positive cells were detected in the primary tumors. Most red signal was not in the cytoplasm of cells associated with nuclei and, therefore, considered nonspecific [insets (a), (c)]. The lungs containing metastases on the other hand [(b), (d)] contained a large number of bona fide luciferase-positive cells corresponding to fusion products. Scale bars on slide scans = 100 μm. Scale bars on 40× inset = 25 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31069316), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Luciferase Antibody (Luci 21 1-107) - BSA Free

Western Blot: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] -

Western Blot: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] - TTFields induce autophagy in glioma cell lines.a U-87 MG & A172 cells were either left untreated or treated with TTFields at the last 24 h, 48 h, or 72 h of culturing. All cultures were plated on the same time, incubated overnight to allow cell attachment, & collected 72 h afterwards. Cells were collected, lysed, & samples were analyzed using immunoblotting for LC3 & GAPDH. Upper panel: representative blots. Lower panel: densitometric quantification of immunoblot signal, showing an average of at least three independent experiments (0.01 < *P < 0.05, **P < 0.01, Student’s t-test). b Paraffin-embedded sections from sham- or TTFields-treated rats were stained with anti-LC3 Ab (green) & DAPI (blue). Representative images are presented. c Quantification of LC3 intensity, presented as fold increase from corresponding control (*P < 0.05, Student’s t-test) Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30341282), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Luciferase Antibody (Luci 21 1-107) - BSA Free

Immunocytochemistry/ Immunofluorescence: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] -

Immunocytochemistry/ Immunofluorescence: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] - TTFields application leads to increased autophagic flux.a Ultra-structural STEM electron microscopy analysis of U-87 MG (upper panel) & A172 (lower panel) cells treated with TTFields for 48 h. Autophagosomes (blue arrows) & autolysosomes (green arrows) are indicated. b U-87 MG & A172 cells were either left untreated or were treated with TTFields for 24–72 h. CQ (20 µM) was added 4 h before cells were collected. Samples were immunoblotted for LC3 & GAPDH. Upper panel: representative blots. Lower panel: densitometric quantification of immunoblot signal, showing an average of at least three independent experiments (0.01 < *P < 0.05, Student’s t-test). c U-87 MG & A172 cells were either left untreated or were treated with TTFields for 48 h. CQ (20 µM) was added at the last 3 h of treatment & cells were fixed & stained with anti-LC3 Ab (green) & DAPI (blue). Upper panel: representative images. Original magnifications: × 40. Lower panel: quantification of LC3 intensity, presented as average intensity per cell (**P < 0.01, Student’s t-test). d U-87 MG cells were either left untreated or were treated with TTFields (48 h) or with vinblastin 25 nM (0.5 h). CQ (20 µM) was added at the last 3 h of treatment & cells were fixed & stained with anti-LC3 (green), LAMP1 (red), & DAPI (blue) (upper panel). Arrowheads indicate the strongest colocalization staining in each cell. Intensity histograms of LAMP1 & LC3 fluorescent signal calculated from the region of interest indicated by the white bar (lower panel). Representative images are shown Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30341282), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
Luciferase Antibody (Luci 21 1-107) - BSA Free

Immunocytochemistry/ Immunofluorescence: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] -

Immunocytochemistry/ Immunofluorescence: Luciferase Antibody (Luci 21 1-107) - BSA Free [NB600-307] - TTFields application leads to increased autophagic flux.a Ultra-structural STEM electron microscopy analysis of U-87 MG (upper panel) & A172 (lower panel) cells treated with TTFields for 48 h. Autophagosomes (blue arrows) & autolysosomes (green arrows) are indicated. b U-87 MG & A172 cells were either left untreated or were treated with TTFields for 24–72 h. CQ (20 µM) was added 4 h before cells were collected. Samples were immunoblotted for LC3 & GAPDH. Upper panel: representative blots. Lower panel: densitometric quantification of immunoblot signal, showing an average of at least three independent experiments (0.01 < *P < 0.05, Student’s t-test). c U-87 MG & A172 cells were either left untreated or were treated with TTFields for 48 h. CQ (20 µM) was added at the last 3 h of treatment & cells were fixed & stained with anti-LC3 Ab (green) & DAPI (blue). Upper panel: representative images. Original magnifications: × 40. Lower panel: quantification of LC3 intensity, presented as average intensity per cell (**P < 0.01, Student’s t-test). d U-87 MG cells were either left untreated or were treated with TTFields (48 h) or with vinblastin 25 nM (0.5 h). CQ (20 µM) was added at the last 3 h of treatment & cells were fixed & stained with anti-LC3 (green), LAMP1 (red), & DAPI (blue) (upper panel). Arrowheads indicate the strongest colocalization staining in each cell. Intensity histograms of LAMP1 & LC3 fluorescent signal calculated from the region of interest indicated by the white bar (lower panel). Representative images are shown Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30341282), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for Luciferase Antibody (Luci 21 1-107) - BSA Free

Application
Recommended Usage

Flow Cytometry

1:200. Use reported by customer review

Immunocytochemistry/ Immunofluorescence

1:100-1:1000

Immunohistochemistry

1:100-1:1000

Immunohistochemistry-Frozen

reported in scientific literature (PMID 31069316)

Immunohistochemistry-Paraffin

1:100-1:1000

Western Blot

1:1000
Application Notes
Western blot has been tested with Drosophila embryos. Purified Luciferase protein and Luciferase expressed in Drosophila adult co-migrate on Western blots with a band seen at ~61 kDa, representing Luciferase.

Reviewed Applications

Read 2 reviews rated 4.5 using NB600-307 in the following applications:

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Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.

Background: Luciferase

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

Alternate Names

LuC, luciferin 4 monooxygenase, Luciferin 4-monooxygenase

Additional Luciferase Products

Product Documents for Luciferase Antibody (Luci 21 1-107) - BSA Free

Certificate of Analysis

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Product Specific Notices for Luciferase Antibody (Luci 21 1-107) - BSA Free

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.

Citations for Luciferase Antibody (Luci 21 1-107) - BSA Free

Customer Reviews for Luciferase Antibody (Luci 21 1-107) - BSA Free (2)

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Customer Images


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  • Name: Anonymous
    Application: Flow Cytometry
    Verified Customer | Posted 12/05/2012
    Luciferase Antibody (Luci 21 1-107) - BSA Free NB600-307
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: cardiomyocytes cell lysate, Sample Amount: 15ug
    Species: Rat
    Verified Customer | Posted 04/22/2010

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Protocols

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FAQs for Luciferase Antibody (Luci 21 1-107) - BSA Free

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  • Q: Hello. Do you have a specific protocol for use of this antibody in FACS analysis? Luciferase Antibody (Luci 21 1-107) 0.1ml. It is uncojugated, so I am wondering if there is a specific protocol for labeling, etc. Thank you

    A:

    The flow protocol used with this antibody NB600-307 was our general flow protocol. Since this antibody is unconjugated, you would want to use a conjugated mouse IgG secondary with this product in order to detect the antibody. We do also have this specific clone available in a directly conjugated PE form (NB600-307PE). If you are interested, I can inquire about a custom conjugate of another fluor or dye if you like. The other option would be for you to directly conjugate the antibody yourself. We do sell several conjugation kits that are very easy to use.

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