Anti-NF-L Polyclonal Antibody 2 is a Rabbit antibody targeting NF-L. Anti-NF-L Polyclonal Antibody 2 can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat

1. 25 μg total protein per lane of various lysates (see on figure) probed with NF-L polyclonal antibody, unconjugated (TMAB-09446) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r. T. for 60 min.
2. Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with NF-L Polyclonal Antibody, Unconjugated (TMAB-09446) at 1: 200 overnight at 4°C. Followed by conjugated Goat Anti-Rabbit IgG antibody (Purple), DAPI (blue) was used to stain the cell nuclei.
3. Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with NF-L Polyclonal Antibody, Unconjugated (TMAB-09446) at 1: 200 overnight at 4°C. Followed by conjugated Goat Anti-Rabbit IgG antibody (Purple), DAPI (blue) was used to stain the cell nuclei.
4. Paraformaldehyde-fixed, paraffin embedded Rat Cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with NF-L Polyclonal Antibody, Unconjugated (TMAB-09446) at 1: 200 overnight at 4°C. Followed by conjugated Goat Anti-Rabbit IgG antibody (Purple), DAPI (blue) was used to stain the cell nuclei.
5. Paraformaldehyde-fixed, paraffin embedded Mouse Cerebellum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with NF-L Polyclonal Antibody, Unconjugated (TMAB-09446) at 1: 200 overnight at 4°C. Followed by conjugated Goat Anti-Rabbit IgG antibody (Purple), DAPI (blue) was used to stain the cell nuclei.
6. Paraformaldehyde-fixed, paraffin embedded Human Left Parietal Lobe; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with NF-L Polyclonal Antibody, Unconjugated (TMAB-09446) at 1: 200 overnight at 4°C. Followed by conjugated Goat Anti-Rabbit IgG antibody (Purple), DAPI (blue) was used to stain the cell nuclei.
7. Paraformaldehyde-fixed, paraffin embedded (mouse spinal cord); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (NF-L) Polyclonal Antibody, Unconjugated (TMAB-09446) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
8. Paraformaldehyde-fixed, paraffin embedded (rat spinal cord); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (NF-L) Polyclonal Antibody, Unconjugated (TMAB-09446) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
9. The U251 (H) cells were fixed with 4% PFA (10 min at r. T.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, the cells then were incubated in 5% BSA to block non-specific protein-protein interactions (30 min at r. T.). Primary Antibody (green): Rabbit Anti-NF-L antibody (TMAB-09446): 1 μg/10^6 cells; Isotype Control (orange): Rabbit IgG. Blank control (black): PBS. Acquisition of 20,000 events was performed.
10. Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (NF-L) Polyclonal Antibody, Unconjugated (TMAB-09446) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
11. Paraformaldehyde-fixed, paraffin embedded (rat cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (NF-L) Polyclonal Antibody, Unconjugated (TMAB-09446) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
12. Paraformaldehyde-fixed, paraffin embedded (human cerebellum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (NF-L) Polyclonal Antibody, Unconjugated (TMAB-09446) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
13. Paraformaldehyde-fixed, paraffin embedded (rat Cerebrum); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (NF-L) Polyclonal Antibody, Unconjugated (TMAB-09446) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

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 NH-L intermedial

Human

NEFL

Neurofilament light polypeptide

Neurofilaments usually contain three intermediate filament proteins: L, M, and H which are involved in the maintenance of neuronal caliber.