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Antibodies

Luciferase Antibody

Product Sizes
25 ul
200-101-C44S-25UL
About this Product
SKU:
200-101-C44S
Additional Names:
goat anti-Luciferase Antibody, Luciferin 4-monooxygenase, Photinus pyralis (Common eastern firefly) (Lampyris pyralis)
Application:
Detection/Quantitation, ELISA, Immunofluorescence, Immunohistochemistry, Western Blot
Buffer:
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride
CE/IVD:
RUO
Clonality:
Polyclonal
Concentration:
1 mg/ml
Extra Details:
Anti luciferase Antibody recognizes luciferase that is commonly used in biological research as a reporter to assess the transcriptional activity in cells that are transfected with a genetic construct containing the luciferase gene under the control of a promoter of interest. Luciferase can also be used to detect the level of cellular ATP in cell viability assays or for kinase activity assays. Additionally proluminescent molecules that are converted to luciferin upon activity of a particular enzyme can be used to detect enzyme activity in coupled or two-step luciferase assays. Such substrates have been used to detect caspase activity and cytochrome P450 activity, among others.
Formulation:
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Host:
Goat
Immunogen:
Luciferase [Photinus pyralis (Firefly)]
Isotype:
IgG
Physical State:
Liquid
Reactivities:
Insect/Arthropod
Shipping Conditions:
Dry Ice
Specificity:
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
Storage Conditions:
-20[o]C diluted. Avoid freeze/thaw cycles., See Manual Avoid freeze/thaw cycles.
Supplier:
Rockland Inc
Type:
Antibody: Polyclonal Antibody
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.
NIH3T3 cells transiently transfected with a luciferase gene. Luciferase-positive cells were detected using Rockland's polyclonal Anti-Luciferase antibody. Cells (25,000/well) were transiently transfected with a pLuc plasmid. After 2 days, the cells were fixed using 4% paraformaldehyde, permeabilized with 0.1% Triton® X-100, and blocked with 1% normal donkey serum. Cells were stained with 20ug/ml Anti-Luciferase in PBS for 2 hours followed by 1:200 dilution of donkey anti-goat IgG-FITC (green) for 1 hour. Cells were mounted using Vectashield with DAPI (blue) and visualized at 200X magnification with a fluorescent microscope.
ELISA Results of Goat Anti-Luciferase Antibody. Each well was coated in duplicate with 1.0 ug of Luciferase [Firefly]. The starting dilution of antibody was 5ug/ml and the X-axis represents the Log10 of a 3-fold dilution. The titer is 1:6,050. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using TMB substrate p/n TMBE-1000.
Western Blot of Goat anti-Luciferase antibody. Lane 1 Marker: Opal Pre-stained ladder (p/n MB-210-0500). Lane 2: HEK293 lysate (p/n W09-000-365). Lane 3: HeLa Lysate (p/n W09-000-364). Lane 4: CHO/K1 Lysate (p/n W07-000-357). Lane 5: E-coli HCP Control (p/n 000-001-J08). Lane 6: J774A.1 Lysate (p/n W10-001-GX3). Lane 7: C2C12 Lysate (p/n W10-001-GL7). Lane 8: Mouse Embryonic Fibroblast Lysate (p/n W10-001-371). Lane 9: NIH/3T3 Lysate (p/n W10-000-358). Lane 10: Mouse Liver Lysate (p/n W10-000-T020). Lane 11: PC-12 Lysate (p/n W12-001-GL9). Lane 12: Rat Brain Lysate (p/n W12-000-T077). Lane 13: Rat Testis Lysate (p/n W12-000-GZ3). Lane 14: Luciferase [Photinus pyralis (Firefly)]. Lane 15 Marker: Opal Pre-stained ladder (p/n MB-210-0500). Load: 10 ug of lysate or 50ng of purified protein per lane. Primary antibody: Luciferase antibody at 1:1,000 overnight at 4C. Secondary antibody: Peroxidase Goat secondary antibody (p/n 605-703-125) at 1:40,000 for 60 min at RT. Blocking Buffer: 1% Casein-TTBS (MB-082) for 30 min at RT. Predicted/Observed size: ~60 kDa for Luciferase.
Western Blot of Goat anti-Luciferase Antibody. Lane 1: Luciferase. Load: 50ng per lane. Primary antibody: Luciferase antibody at 1:1000 overnight at 4°C. Secondary antibody: Peroxidase goat secondary antibody (p/n 611-103-122) at 1:40,000 for 30 min at RT. Block: (p/n MB-070) for 30 min at RT. Predicted/Observed size: ~60 kDa for Luciferase. Other band(s): None.