Antibody Type: Recombinant Monoclonal

Application: ELISA, WB, IP

Reactivity: Human

Rabbit IgG

5R792

Human

1. Western Blot
-Positive WB detected in: JK whole cell lysate(20µg), K562 whole cell lysate(20µg), SY5Y whole cell lysate(20µg), HepG2 whole cell lysate(20µg), COLO205 whole cell lysate(20µg)
-All lanes: CDK6 antibody at 1:1000
-Secondary: Goat polyclonal to rabbit IgG at 1/40000 dilution
-Predicted band size: 37 kDa
-Observed band size: 37 kDa
-Exposure time: 120s
2. Immunofluorescence staining of U251 cell with TMAH-00255 at 1:10, 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).
3. Immunofluorescence staining of U251 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).
4. Immunofluorescence staining of Hela cell with TMAH-00255 at 1:10, 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).
5. Immunofluorescence staining of Hela 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).
6. IHC image of TMAH-00255 diluted at 1:50 and staining in paraffin-embedded human tonsil 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 polymer IgG labeled by HRP and visualized using 0.05% DAB. Secondary antibody only control: uses 1% BSA instead of primary antibody
7. IHC image of TMAH-00255 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
8. Immunoprecipitating CDK6 in K562whole cell lysate
-Lane 1: TMAH-00255(3µg)+ K562 whole cell lysate(220µg)
-Lane 2: K562 whole cell lysate(30µg)
-Lane 3:Rabbit control IgG instead of TMAH-00563 in K562 whole cell lysate
For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/40000)

ELISA, WB, IP

WB:1:500-1:5000; IP:1:200-1:1000.

Monoclonal

Unconjugated

A synthetic peptide: Human CDK6

Human

1021

Epigenetics and Nuclear Signaling, Cancer, Cell biology