Anti-Cystathionase/CTH antibody [2E12-1C10]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystathionase/CTH antibody [2E12-1C10] (AB54573)

Cystathionase/CTH antibody (ab54573) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human colon.

This image was generated using the ascites version of the product.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Cystathionase/CTH antibody [2E12-1C10] (AB54573)

ICC/IF image of ab54573 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab54573, 5μg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.
This image was generated using the ascites version of the product.

• Flow Cyt

Unknown

Flow Cytometry - Anti-Cystathionase/CTH antibody [2E12-1C10] (AB54573)

Overlay histogram showing K562 cells stained with ab54573 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab54573, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in K562 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

This image was generated using the ascites version of the product.

• WB

Lab

Western blot - Anti-Cystathionase/CTH antibody [2E12-1C10] (AB54573)

Western blot : Anti-CTH antibody [2E12-1C10] (ab54573) staining at 1/1000 dilution, shown in green; Rabbit anti-ACTN4 [EPR2533(2)] (ab108198) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab54573 was shown to bind specifically to CTH. A band was observed at 42 kDa in wild-type A549 cell lysates with no signal observed at this size in CTH knockout cell line. To generate this image, wild-type and CTH knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Cystathionase/CTH antibody [2E12-1C10] (ab54573) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

CTH knockout A549 cell lysate at 20 µg

Lane 3:

K562 cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 42 kDa

false

• WB

Unknown

Western blot - Anti-Cystathionase/CTH antibody [2E12-1C10] (AB54573)

Cystathionase/CTH antibody (ab54573) at 1ug/lane + K-562 cell lysate at 25ug/lane.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-Cystathionase/CTH antibody [2E12-1C10] (ab54573)

Predicted band size: 45 kDa

false