Antibody Type: Recombinant Rabbit Monoclonal

Application: ELISA, WB, IHC, IF, FCM, IP

Reactivity: Human

Rabbit IgG

1L745

Human

1. Western Blot
-Positive WB detected in: Jurkat whole cell lysate, K562 whole cell lysate, Hela whole cell lysate, Raji whole cell lysate, HepG2 whole cell lysate
-All lanes: FUBP1 antibody at 1:2000
-Secondary: Goat polyclonal to rabbit IgG at 1/50000 dilution
-Predicted band size: 68, 69 kDa
-Observed band size: 69 kDa
2. IHC image of TMAH-00464 diluted at 1:100 and staining in paraffin-embedded human brain tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB.
3. IHC image of TMAH-00464 diluted at 1:100 and staining in paraffin-embedded human glioma cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB.
4. Immunofluorescence staining of Hela Cells with TMAH-00464 at 1:50, counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeated by 0.2% TritonX-100, and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. Nuclear DNA was labeled in blue with DAPI. The secondary antibody was FITC-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L).
5. Overlay histogram showing Jurkat cells stained with TMAH-00464 (red line) at 1:50. The cells were fixed with 70% Ethylalcohol (18h) and then incubated in 10% normal goat serum to block non-specific protein-protein interactions followedby the antibody (1µg/1*10^6 cells) for 1 h at 4°C.The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 30min at 4°C. Control antibody (green line) was Rabbit IgG (1µg/1*10^6 cells) used under the same conditions. Acquisition of >10,000 events was performed.
6. Immunoprecipitating FUBP1 in Jurkat whole cell lysate
-Lane 1: Rabbit control IgG instead of TMAH-00464 in Jurkat whole cell lysate.
For western blotting,a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
-Lane 2: TMAH-00464(2µg)+ Jurkat whole cell lysate(500µg)
-Lane 3: Jurkat whole cell lysate (10µg)

ELISA, WB, IHC, IF, FCM, IP

WB:1:500-1:5000; IHC:1:50-1:200; IF:1:20-1:200; FCM:1:20-1:200; IP:1:200-1:1000.

Monoclonal

Unconjugated

A synthetic peptide: Human FUBP1

Human

8880

Epigenetics and Nuclear Signaling