Anti-Oct-3/Oct-4 Polyclonal Antibody is a Rabbit antibody targeting Oct-3/Oct-4. Anti-Oct-3/Oct-4 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

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

1. Tissue/cell: mouse tooth germ 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-Oct-3/Oct-4 Polyclonal Antibody, Unconjugated (TMAB-09744) 1:100, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
2. Paraformaldehyde-fixed, paraffin embedded (rat placenta); 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 (Oct-3) Polyclonal Antibody, Unconjugated (TMAB-09744) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (mouse placenta); 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 (Oct-3) Polyclonal Antibody, Unconjugated (TMAB-09744) at 1: 200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
4. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-Oct-3 antibody (TMAB-09744)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
5. Sample:
Embryo (Mouse) Lysate at 40 μg
Testis (Mouse) Lysate at 40 μg
NIH/3T3 (Mouse) Cell Lysate at 40 μg
Hela (Human) Cell Lysate at 40 μg
Rsc96 (Rat) Cell Lysate at 40 μg
293T (Human) Cell Lysate at 40 μg
SP2/0 (Mouse) Cell Lysate at 40 μg
HL-60 (Human) Cell Lysate at 40 μg
Primary: Anti-Oct-3/Oct-4 (TMAB-09744) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 39 kD
Observed band size: 39 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 OCT4

Human

POU5F1

POU domain, class 5, transcription factor 1

Transcription factor that binds to the octamer motif (5'-ATTTGCAT-3'). Forms a trimeric complex with SOX2 on DNA and controls the expression of a number of genes involved in embryonic development such as YES1, FGF4, UTF1 and ZFP206. Critical for early embryogenesis and for embryonic stem cell pluripotency.