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

IgG

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

1. Tissue/cell: human liver 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-Insulin Receptor/CD220 Polyclonal Antibody, Unconjugated (TMAB-00974) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Tissue/cell: rat liver 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-Insulin Receptor/CD220 Polyclonal Antibody, Unconjugated (TMAB-00974) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Blank control: Raji (blue). Primary Antibody: Rabbit Anti-Insulin Receptor alpha antibody (TMAB-00974), 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. Primary antibody (TMAB-00974, 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.
4. Sample:
Lane 1: HepG2 (Human) Cell Lysate at 30 μg
Lane 2: 293T (Human) Cell Lysate at 30 μg
Lane 3: LOVO (Human) Cell Lysate at 30 μg
Lane 4: Molt-4 (Human) Cell Lysate at 30 μg
Lane 5: SW480 (Human) Cell Lysate at 30 μg
Lane 6: K562 (Human) Cell Lysate at 30 μg
Primary: Anti-Insulin Receptor (TMAB-00974) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 120 kDa
Observed band size: 120 kDa
5. Sample:
Lane 1: MCF-7 (Human) Cell Lysate at 30 μg
Lane 2: A549 (Human) Cell Lysate at 30 μg
Lane 3: 293T (Human) Cell Lysate at 30 μg
Lane 4: Hela (Human) Cell Lysate at 30 μg
Lane 5: Liver (Rat) Lysate at 40 μg
Lane 6: LympHnode (Mouse) Lysate at 40 μg
Lane 7: MOLT-4 (Human) Cell Lysate at 30 μg
Lane 8: NIH/3T3 (Mouse) Cell Lysate at 30 μg
Primary: Anti-Insulin Receptor (TMAB-00974) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 120 kDa
Observed band size: 120 kDa

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

Polyclonal

KLH conjugated synthetic peptide: human Insulin Receptor

Human

Insulin and insulin-like,Diabetes associated,Diabetes,Heart disease,Metabolism,Insulin / Insulin-like,Receptor Tyrosine Kinases