All lanes : Anti-Citrate synthetase antibody (ab96600) at 1/1000 dilution

Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 2 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : HRP-conjugated anti-rabbit IgG

Predicted band size: 52 kDa

10% SDS-PAGE.

Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Citrate synthetase (green) with ab96600 at a dilution of 1/200.

Cells were fixed in 4% paraformaldehyde at room temperature for 15 minutes. Alpha Tubulin (red) was stained with ab184613 at a dilution of 1/1000. Blue: Hoechst 33342 staining.

Anti-Citrate synthetase antibody (ab96600) at 1/1000 dilution + Rat muscle tissue extracts at 50 µg

Predicted band size: 52 kDa

10% SDS-PAGE.

Anti-Citrate synthetase antibody (ab96600) at 1/1000 dilution + Rat brain tissue extracts at 50 µg

Secondary
HRP-conjugated anti-rabbit IgG

Predicted band size: 52 kDa

10% SDS-PAGE.

Anti-Citrate synthetase antibody (ab96600) at 1/1000 dilution + Mouse brain tissue extracts at 50 µg

Secondary
HRP-conjugated anti-rabbit IgG

Predicted band size: 52 kDa

10% SDS-PAGE.

Immunocytochemistry/Immunofluorescence analysis of U-2 OS (Human bone osteosarcoma epithelial cell line) cells labelling Citrate synthetase (green) with ab96600 at a dilution of 1/500.

Cells were fixed in 4% paraformaldehyde at room temperature for 15 minutes. Alpha Tubulin (red) was stained with ab184613 at a dilution of 1/1000. Blue: Hoechst 33342 staining.

IHC image of ab96600 staining in human colon formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab96600, 5 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Immunoprecipitation of Citrate synthetase from HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate using 5 μg of ab96600.

Western blot analysis was performed using ab96600.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse intestine tissue labeling Citrate synthetase with ab96600 at a dilution of 1/500.

Antigen Retrieval: EDTA based buffer, 15 mins.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse muscle tissue labeling Citrate synthetase with ab96600 at a dilution of 1/500.

Antigen Retrieval: Citrate buffer, pH 6.0, 15 mins.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of NCI-N87 xenograft tissue labeling Citrate synthetase with ab96600 at a dilution of 1/500.

Antigen Retrieval: EDTA based buffer, 15 mins.

All lanes : Anti-Citrate synthetase antibody (ab96600) at 1 µg/ml

Lane 1 : Marker
Lane 2 : Zebrafish liver homogenate (20ug)
Lane 3 : Zebrafish skeletal muscle homogenate (20ug)
Lane 4 : Mouse liver mitochondrial homogenate (20ug)

Secondary
All lanes : Goat polyclonal to Rabbit IgG – H&L – Pre-Adsorbed (HRP) at 1/6000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 52 kDa
Observed band size: 47 kDa why is the actual band size different from the predicted?


Exposure time: 5 minutes