NMNAT1 antibody | knockout validation | Cell Signaling 98354

This is a knockout-validated antibody summary, based on the publication "Nuclear NAD + homeostasis governed by NMNAT1 prevents apoptosis of acute myeloid leukemia stem cells", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

NMNAT1 antibody | knockout validation | Cell Signaling 98354 figure 1
Figure 1. Immunoblot of NMNAT1 in MOLM13 cells at day 4 after deleting the gene. From [1].
Antibody information

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: NMNAT1

Catalog number: 98354

Summary: Rabbit monoclonal antibody against a synthetic peptide corresponding to residues surrounding Ala268 of human NMNAT1 protein. Reacts with human by western blot and immunoprecipitation.

Validation Method

Western blot

Sample

Control and NMNAT1 KO MOLM13 cells.

Blocking agent

5% (w/v) nonfat milk in 1× PBST (PBS with 0.1% Tween-20) for 1 hour at room temperature.

Primary incubation

1:1,000 dilution overnight at 4°C.

Secondary incubation

1:5,000-10,000 dilution anti-rabbit horseradish peroxidase–conjugated secondary antibodies (Cell Signaling Technology).

Detection

SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo Fisher Scientific, catalog no. 34580) and a KODAK X-OMAT 2000 Processor (Kodak).

References
  1. Shi X, Jiang Y, Kitano A, Hu T, Murdaugh R, Li Y, et al. Nuclear NAD+ homeostasis governed by NMNAT1 prevents apoptosis of acute myeloid leukemia stem cells. Sci Adv. 2021;7: pubmed publisher