重组Anti-DGCR8抗体[EPR18757]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DGCR8 antibody [EPR18757] (AB191875)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling DGCR8 with ab191875 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on HeLa cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -
-ve control 1 : ab191875 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DGCR8 antibody [EPR18757] (AB191875)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling DGCR8 with ab191875 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear and weakly cytoplasmic staining on Jurkat cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -
-ve control 1 : ab191875 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-DGCR8 antibody [EPR18757] (AB191875)

Intracellular Flow Cytometry analysis of Jurkat (human acute T cell leukemia) labelling DGCR8 with purified ab191875 at 1/1000 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Alexa Fluor^® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

• IP

Supplier Data

Immunoprecipitation - Anti-DGCR8 antibody [EPR18757] (AB191875)

DGCR8 was immunoprecipitated from 1mg of HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate with ab191875 at 1/60 dilution.

Western blot was performed from the immunoprecipitate using ab191875 at 1/1000 dilution.

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : HEK-293 whole cell lysate 10ug (Input).

Lane 2 : ab191875 IP in HEK-293 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab191875 in HEK-293 whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-DGCR8 antibody [EPR18757] (ab191875)

Predicted band size: 86 kDa

false

• WB

Lab

Western blot - Anti-DGCR8 antibody [EPR18757] (AB191875)

Western blot : Anti-DGCR8 antibody [EPR18757] (ab191875) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab191875 was shown to bind specifically to DGCR8. A band was observed at 100 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in DGCR8 knockout cell line. To generate this image, wild-type and DGCR8 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

Western blot - Human DGCR8 knockout HCT116 cell line (<a href='/products/cell-lines/human-dgcr8-knockout-hct116-cell-line-ab287366'>ab287366</a>)

Lane 2:

DGCR8 knockout HCT 116 cell lysate at 20 µg

Lane 3:

Wild-type A549 ab288558 cell lysate at 20 µg

Lane 4:

DGCR8 knockout A549 <a href='/products/cell-lines/human-dgcr8-knockout-a549-cell-line-ab287368'>ab287368</a> cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 100 kDa

false

• WB

Supplier Data

Western blot - Anti-DGCR8 antibody [EPR18757] (AB191875)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875) at 1/1000 dilution

All lanes:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

Predicted band size: 86 kDa

Observed band size: 100 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-DGCR8 antibody [EPR18757] (AB191875)

Western blot : Anti-DGCR8 antibody [EPR18757] (ab191875) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab191875 was shown to bind specifically to DGCR8. A band was observed at 90-100 kDa in wild-type A549 cell lysates with no signal observed at this size in DGCR8 knockout cell line. To generate this image, wild-type and DGCR8 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 5 % 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-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

DGCR8 knockout A549 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

PC-3 cell lysate at 20 µg

Secondary

All lanes:

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

false

• WB

Supplier Data

Western blot - Anti-DGCR8 antibody [EPR18757] (AB191875)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875) at 1/1000 dilution

Lane 1:

HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 3:

WEHI-3 (Mouse leukemia cell line) whole cell lysate at 20 µg

Lane 4:

Neuro-2a (Mouse neuroblastoma cells) whole cell lysate at 20 µg

Lane 5:

Mouse testis lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 86 kDa

Observed band size: 100 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-DGCR8 antibody [EPR18757] (AB191875)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Rat brain lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 86 kDa

Observed band size: 100 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-DGCR8 antibody [EPR18757] (AB191875)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-DGCR8 antibody [EPR18757] (ab191875) at 1/1000 dilution

Lane 1:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 10 µg

Lane 2:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 86 kDa

Observed band size: 100 kDa

false

Exposure time: 5s