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

IgG

Mouse,Rat (predicted:Cow,Rabbit)

1. Tissue/cell: rat colon tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-L-Selectin Polyclonal Antibody, Unconjugated (TMAB-01084) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Tissue/cell: rat colitis tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-L-Selectin Polyclonal Antibody, Unconjugated (TMAB-01084) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated used at 1:200 dilution for 40 minutes at 37°C. DAPI (5 μg/ml,blue) was used to stain the cell nucleus.
3. Tissue/cell: mouse kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-L-Selectin Polyclonal Antibody, Unconjugated (TMAB-01084) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
4. Blank control: mouse spleen cells (blue). Primary Antibody: Rabbit Anti-CD62L antibody (TMAB-01084), 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). Primary antibody (TMAB-01084, 1 μg/1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 10% 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 at 1/200 dilution for 30 min on ice.
5. Sample:
LympHnode (Mouse) Lysate at 40 μg
LympHnode (Rat) Lysate at 40 μg
Primary: Anti-CD62L (TMAB-01084) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 37 kDa
Observed band size: 43 kDa

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: mouse CD62L

Mouse

Blood,Regulatory T Cells,CD,T Lymphocytic Lineage,Neutrophil Lineage