重组Anti-Drosha抗体[EPR12794]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Drosha antibody [EPR12794] (AB183732)

Immunohistochemical analysis of paraffin-embedded Human lung adenocarcinoma tissue labeling Drosha with ab183732 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• WB

Unknown

Western blot - Anti-Drosha antibody [EPR12794] (AB183732)

Lanes 1-3 : Merged signal (red and green). Green - ab183732 observed at 159 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab183732 Anti-Drosha antibody [EPR12794] was shown to specifically react with Drosha in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266217 (knockout cell lysate ab257171) was used. Wild-type and Drosha knockout samples were subjected to SDS-PAGE. ab183732 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Drosha antibody [EPR12794] (ab183732) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

DROSHA knockout HEK293T cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Predicted band size: 159 kDa

false

• WB

Lab

Western blot - Anti-Drosha antibody [EPR12794] (AB183732)

Lanes 1-3 : Merged signal (red and green). Green - ab183732 observed at 159 kDa. Red - loading control, ab8245 observed at 37 kDa. ab183732 Anti-Drosha antibody [EPR12794] was shown to specifically react with Drosha in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266217 (knockout cell lysate ab257171) was used. Wild-type and Drosha knockout samples were subjected to SDS-PAGE. ab183732 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Drosha antibody [EPR12794] (ab183732)

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

DROSHA knockout HEK-293T cell lysate, at 20 µg

Lane 2:

Western blot - Human DROSHA knockout A549 cell line (<a href='/products/cell-lines/human-drosha-knockout-a549-cell-line-ab287377'>ab287377</a>)

false

• WB

Lab

Western blot - Anti-Drosha antibody [EPR12794] (AB183732)

Lanes 1-3 : Merged signal (red and green). Green - ab183732 observed at 159 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab183732 Anti-Drosha antibody [EPR12794] was shown to specifically react with Drosha in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266217 (knockout cell lysate ab257171) was used. Wild-type and Drosha knockout samples were subjected to SDS-PAGE. ab183732 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Drosha antibody [EPR12794] (ab183732) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

DROSHA knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human DROSHA knockout HEK-293T cell line (<a href='/products/cell-lines/human-drosha-knockout-hek-293t-cell-line-ab266217'>ab266217</a>)

Lane 3:

HeLa cell lysate at 20 µg

Predicted band size: 159 kDa

Observed band size: 159 kDa

false

• WB

Supplier Data

Western blot - Anti-Drosha antibody [EPR12794] (AB183732)

All lanes:

Western blot - Anti-Drosha antibody [EPR12794] (ab183732) at 1/50000 dilution

Lane 1:

Jurkat cell lysate at 10 µg

Lane 2:

293 cell lysate at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 159 kDa

Observed band size: 159 kDa

false

• WB

CiteAb

Western blot - Anti-Drosha antibody [EPR12794] (AB183732)

Drosha western blot using anti-Drosha antibody [EPR12794] ab183732. Publication image and figure legend from Mantel, P. Y., Hjelmqvist, D., et al., 2016, Nat Commun, PubMed 27721445.

ab183732 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab183732 please see the product overview.

Detection of human Ago2 in RBCs and iRBCs.(a) Expression of RNAi machinery during RBC development. Cell lysates from different developmental stages of erythropoiesis were prepared (day 11 : basophilic and polychromatic erythroblasts, day 18 : orthochromatic erythroblasts and reticulocytes, day 21 : reticulocytes and normocytes; representative images are shown). Ago2 is detectable in mature RBCs while dicer, drosha (and human histone H3) are not present. Stomatin is present in all preparations. (b) IFA of Ago2 in RBCs and iRBCs. Ago2 is localized to the RBC periphery in uninfected and infected RBCs, however, labelling is reduced in later parasite stages. Notably, ring stage parasites also show Ago2 accumulation in the parasite. Scale bar; 1 μm. (c) Flow cytometry analysis of Ago2 labelling in RBCs and iRBCs. Uninfected RBCs and iRBCs were gated based on SYBR staining (nuclear content, n) and Ago2 labelling. Young iRBCs (rings, and trophozoites with n=1) show the highest Ago2 labelling, confirming IFA data. (d) Sequential fractionation of purified infected parasites analysed by western blot. iRBC samples were collected at four time points post invasion, separated from uninfected RBCs and fractionated. The host cytosol was released with 1 HU of tetanolysin (TTL) and subsequently the PV contents were released with 0.035% of saponin (SAP). The pellet contains parasite material and host membranes while the TTL supernatant contains RBC cytosol and the SAP supernatant contains soluble parasitophorous vacuole material. The parasitophorous vacuole membrane marker PfExp-1 and parasite nuclear histone H3 are detectable in the pellet fraction across the asexual parasite cycle. Ago2 is present in cytosol (SAP, TTL) across the cycle but only present in the pellet fraction in early parasite stages, supporting the dynamic distribution observed by IFA. Host stomatin is present in cytosol (SAp, TTL) and membranes (pellet) throughout the cycle. Data are representative of three independent experiments. pi : post invasion.

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