Anti-NME1/NM23A Polyclonal Antibody is a Rabbit antibody targeting NME1/NM23A. Anti-NME1/NM23A Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Dog,Pig,Cow,Horse,Rabbit)

1. Tissue/cell: mouse heart tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-NME1/Nm23-H1/NDKA Polyclonal Antibody, Unconjugated (TMAB-09532) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
2. Blank control: RSC96 (blue). Primary Antibody: Rabbit Anti-NME1 antibody (TMAB-09532), Dilution: 1 μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit IgG (orange), used under the same conditions); Secondary Antibody: Goat anti-rabbit IgG-PE (white blue), Dilution: 1: 200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice. Antibody (TMAB-09532, 1 μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA+1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody of TMAB-09532 at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.
3. Blank control (blue line): A549 (blue).
Primary Antibody (green line): Rabbit Anti-NME1 antibody (TMAB-09532)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 μg /test.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min), then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
4. Sample: Hela (Human) Cell Lysate at 30 μg
Primary: Anti-NM23A (TMAB-09532) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 17 kD
Observed band size: 18 kD

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human Nm23-H1

Human

NME1

Nucleoside diphosphate kinase A

Major role in the synthesis of nucleoside triphosphates other than ATP. Possesses nucleoside-diphosphate kinase, serine/threonine-specific protein kinase, geranyl and farnesyl pyrophosphate kinase, histidine protein kinase and 3'-5' exonuclease activities. Involved in cell proliferation, differentiation and development, signal transduction, G protein-coupled receptor endocytosis, and gene expression. Required for neural development including neural patterning and cell fate determination.