Antibody Type: Recombinant Rabbit Monoclonal

Application: ELISA, WB, IF, FCM

Reactivity: Human, Mouse, Rat

Rabbit IgG

2E46

Human, Mouse, Rat

1. Western Blot
-Positive WB detected in: U87 whole cell lysate, MCF-7 whole cell lysate, PC3 whole cell lysate, Hela whole cell lysate, SH-SY5Y whole cell lysate, THP-1 whole cell lysate, A549 whole cell lysate, Mouse brain tissue, Rat brain tissue
-All lanes: YWHAB antibody at 1:1000
-Secondary: Goat polyclonal to rabbit IgG at 1/50000 dilution
-Predicted band size: 29, 28 kDa
-Observed band size: 25-35 kDa
2. Overlay Peak curve showing Hela cells stained with TMAH-01258 (red line) at 1:100. The cells were fixed in 4% formaldehyde (15min) and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/1*10^6 cells) for 45min at 4°C. The secondary antibody used was FITC-conjugated Goat Anti-rabbit IgG(H+L) at 1:200 dilution for 35min at 4°C.Control antibody (green line) was rabbit IgG (1ug/1*10^6 cells) used under the same conditions. Acquisition of >10,000 events was performed.
3. IHC image of TMAH-01258 diluted at 1:50 and staining in paraffin-embedded human lung 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 polymer IgG labeled by HRP and visualized using 0.05% DAB. Secondary antibody only control: uses 1% BSA instead of primary antibody
4. IHC image of TMAH-01258 diluted at 1:50 and staining in paraffin-embedded human colorectal 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 polymer IgG labeled by HRP and visualized using 0.05% DAB. Secondary antibody only control: uses 1% BSA instead of primary antibody
5. IHC image of TMAH-01258 diluted at 1:50 and staining in paraffin-embedded human breast 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 polymer IgG labeled by HRP and visualized using 0.05% DAB. Secondary antibody only control: uses 1% BSA instead of primary antibody
6. Immunofluorescence staining of A431 cell with TMAH-01258 at 1:20, counter-stained with DAPI. The cells were fixed in 4% formaldehyde and and permeated by 0.2% TritonX-100 for 15 min. Then 10% normal goat serum to block non-specific protein-protein interactions. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L).
7. Immunofluorescence staining of A431 cell with 5% goat serum, counter-stained with DAPI. The cells were fixed in 4% formaldehyde and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L).

ELISA, WB, IF, FCM

WB:1:500-1:2000; IF:1:50-1:200; FCM:1:50-1:200.

Monoclonal

Unconjugated

A synthetic peptide: Human YWHAB

Human

7529

Neuroscience, Cell biology, Signal transduction, Stem cells