Antibody Type: Rabbit Polyclonal

Application: WB,IHC-P,IHC-Fr,IF,FCM

Reactivity: Human,Mouse,Rat (predicted:Chicken,Dog,Pig,Cow,Sheep)

IgG

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

1. Tissue/cell: TMAB-01401 human 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-phospho-CREB-1 (Ser133) Polyclonal Antibody, Unconjugated (TMAB-01401) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Tissue/cell: rat brain 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-phospho-CREB-1 (Ser133) Polyclonal Antibody, Unconjugated (TMAB-01401) 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. Blank control: RSC96 (blue), the cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with ice-cold 90% methanol for 30 min on ice.
Isotype Control Antibody: Rabbit Igg (orange);
Secondary Antibody: Goat anti-rabbit IgG-Pe (white blue),
Dilution: 1:200 in 1 X PBS containing 0.5% BSA;
Primary Antibody Dilution: 1 μg in 100 μL 1X PBS containing 0.5% BSA (green).
4. P-CREB1 was immunoprecipitated from mouse kidney tissue with TMAB-01401 at 1/150 dilution. Western blot was performed from the immunoprecipitate using protein A/G beads. HRP Conjugated Mouse anti-Rabbit IgG (Light Chain specific) was used as secondary antibody at 1:5000 dilution.
Lane 1: mouse kidney tissue lysate 10 µg (Input).
Lane 2: TMAB-01401 IP in mouse kidney tissue lysate.
Lane 3: native rabbit IgG IP in mouse kidney tissue lysate (negative control).
Secondary
All
Lanes: Mouse anti-Rabbit IgG (Light Chain specific), HRP Conjugated, 1:5000
5. Sample:
Lane 1: Cerebrum (Mouse) Lysate at 40 μg
Lane 2: Cerebellum (Mouse) Lysate at 40 μg
Lane 3: Kidney (Mouse) Lysate at 40 μg
Lane 4: Huvec (Human) Cell Lysate at 30 μg
Primary: Anti-phospho-CREB-1 (Ser133) (TMAB-01401) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 43 kDa
Observed band size: 45 kDa

WB,IHC-P,IHC-Fr,IF,FCM

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

Polyclonal

KLH conjugated Synthesised phosphopeptide: human CREB-1 around the phosphorylation site of Ser133

Human

1385

HLH / Leucine Zipper,Transcription Factors,TLR Signaling