Hippocampi were dissected on ice and homogenized in lysis buffer (NaCl 150 mM, Tris 10 mM, 1% Triton X-100, 0.5% nonidet p-40) with protease inhibitor cocktail (EMD Milipore). Fifty μg of proteins were gel separated, transferred to immobilon PVDF-FL membrane. Membranes were incubated in Odyssey blocking buffer (Licor Biosciences) for an hour at room temperature. This was followed by incubation of membranes with primary antibody overnight at 4 °C (anti-PAR (10H), 1:1000 Enzo; anti-Nampt (PBEF), 1:1000 abcam; anti-Nmnat2, 1:1000 abcam, anti-β-actin at 1:10,000; Cell Signaling). After washing the membranes in PBS with 0.1% tween-20, they were incubated with the appropriate infrared fluorophore conjugated secondary antibody (Licor) for 30 min at room temperature. The membranes were scanned and the bands were quantified with Odyssey infrared image system (Licor). The β-actin signal was used for normalization of the data detected from bands of the proteins of interest.
Anti nmnat2
Manufactured by Abcam
Anti-Nmnat2 is a polyclonal antibody that recognizes the Nmnat2 protein. Nmnat2 is an enzyme involved in the biosynthesis of NAD+, a coenzyme important for cellular metabolism and energy production. This antibody can be used to detect and study the Nmnat2 protein in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Odyssey infrared image system, by LI COR (1 mentions)
Odyssey blocking buffer, by LI COR (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Anti par 10h, by Enzo Life Sciences (1 mentions)
Anti parp 1, by Santa Cruz Biotechnology (1 mentions)
Anti cleaved caspase 3, by Santa Cruz Biotechnology (1 mentions)
Anti acetyl p53, by Cell Signaling Technology (1 mentions)
Anti sirt1, by Cell Signaling Technology (1 mentions)
Hrp anti mouse, by Thermo Fisher Scientific (1 mentions)
Anti padpr, by Santa Cruz Biotechnology (1 mentions)
Anti tubulin, by Abcam (1 mentions)
P53 do 1, by Santa Cruz Biotechnology (1 mentions)
Anti caspase 3, by Cell Signaling Technology (1 mentions)
Anti β actin, by Merck Group (1 mentions)
2 protocols using anti nmnat2
1
Western Blot Analysis of Hippocampal Proteins
2
Comprehensive Antibody Protocol for Fly Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following commercially available antibodies were used: anti-Repo (1:250, DSHB, 8D12), anti-Caspase-3 (1:250 for Immunocytochemistry of fly brain, 1:1000 for Western blot analysis, Cell Signaling Technology, 9665), anti-Cleaved Caspase-3 (1:1000, Santa Cruz, 9661), anti-H2AvD (1:50, Rockland, 600-401-914), anti-p53(E-5) (1:50, Santa Cruz, sc-74573), p53(DO-1) (1:1000, Santa Cruz, sc-126), anti-Drosophila Nmnat (1:3000), anti-NMNAT1 (1:1000, Abcam, ab45548), anti-NMNAT1 (1:1000, Santa Cruz, 271557), anti-NMNAT2 (1:500, Abcam, ab56980), anti-PARP1 (1:1000, Santa Cruz, sc-8007), anti-pADPr (1:1000, Santa Cruz, sc-56198), anti-SIRT1 (1:1000, Cell Signaling Technology, 2492), anti-acetyl-p53 (1:1000, Cell Signaling Technology, 2525), anti-β-actin (1:10,000, Sigma-Aldrich, A1978), and anti-tubulin (1:300, Abcam, ab15246). The secondary antibodies conjugated to Alexa 488/546/647 (1:250, Invitrogen), or near-infrared (IR) dye 700/800 (1:5000, LI-COR Biosciences). HRP-anti-mouse and HRP-anti-rabbit (1:5000, Thermo Fisher Scientific).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!