Anti-Firefly Luciferase抗体[Luci17]
参阅全部 Firefly Luciferase 一抗
小鼠单克隆抗体[Luci17] to Firefly Luciferase
Mouse
适用于: WBmore details
与反应: Firefly
Recombinant fragment corresponding to Firefly Firefly Luciferase. Hybridoma produced by the fusion of splenocytes from mice immunized with luciferase protein isolated from Photinus pyralis.
Database link: P08659
WB: Purified luciferase protein
Analysis of gene expression is most commonly assayed by transient transfection. Systems are generally based on the use of fusion genes which are inserted into cells, and the gene expression is assayed within 48 hours after introduction of DNA. Usually the fusion consists of the promoter binding site or enhancer sequence under study which is attached to a reporter gene. The amount of the reporter protein synthesized under the experimental conditions, is presumed to reflect the ability of the sequences studied to direct or promote transcription. Several enzymes are commonly used as reporter proteins, among them are chloramphenicol acetyl transferase (CAT), -galactosidase, human growth hormone (hGH) and luciferase. Luciferase has become one of the widely used reporter enzymes. The enzyme catalyzes a bioluminescent reaction which requires the substrate luciferin as well as Mg+2 and ATP. Mixing these reagents with the cell extract containing luciferase, results in a flash of light that decays rapidly. This light can be detected by a luminometer. The total light emission is proportional to the luciferase activity of the sample. The use of an antibody to detect luciferase can provide an alternative detection assay which directly detects luciferase protein levels, and thus has the advantage that it does not require luciferase activity and is not dependent on rapid kinetics. Moreover, antibodies can detect the luciferase enzyme expression in situ, providing a means to study the localized signal sequences using luciferase as a reporter gene. Reacts with Luciferase (Firefly) Protein.
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Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
pH: 7.20
Preservative: 0.08% Sodium azide
Constituent: PBS
浓度
100 µg 浓度为 1 mg/ml
Protein A/G purified
单克隆
Luci17
IgG1
unknown
Abpromise™承诺保证使用ab16466于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | (2) | Use at an assay dependent concentration. |
ec 1.13.12.7 antibody
Firefly antibody
Luciferase antibody
Luciferin 4 monooxygenase antibody
![Western blot - Anti-Firefly Luciferase antibody [Luci17] (ab16466)](https://www.abcam.cn/ps/products/16/ab16466/Images/ab16466-278059-anti-firefly-luciferase-antibody-luci17-western-blot.jpg)
Western blot - Anti-Firefly Luciferase antibody [Luci17] (ab16466)
All lanes : Anti-Firefly Luciferase antibody [Luci17] (ab16466) at 1 µg/ml
Lane 1 : Non-transfected 293 whole cell lysate
Lane 2 : Firefly Luciferase transfected 293 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : NIH3T3 whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/10000 dilution
Performed under reducing conditions.
Observed band size: 65 kDawhy is the actual band size different from the predicted?
Lanes 1 - 4: Merged signal (red and green). Green – ab16466 observed at 65 kDa. Red - loading control, ab181602, observed at 37 kDa.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab16466 and ab181602 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at a 1:10000 dilution for 1hr at room temperature and then imaged.
