Anti-Dicer antibody [EPR24104-105] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dicer antibody [EPR24104-105] - BSA and Azide free (AB280957)

This data was developed using ab259327, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human testis tissue labelling Dicer with ab259327 at 1/100 dilution (5.05 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human testis (PMID : 17573847). The section was incubated with ab259327 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dicer antibody [EPR24104-105] - BSA and Azide free (AB280957)

This data was developed using ab259327, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling Dicer with ab259327 at 1/100 dilution (5.05 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse testis. The section was incubated with ab259327 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dicer antibody [EPR24104-105] - BSA and Azide free (AB280957)

This data was developed using ab259327, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labelling Dicer with ab259327 at 1/100 dilution (5.05 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat testis. The section was incubated with ab259327 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Dicer antibody [EPR24104-105] - BSA and Azide free (AB280957)

This data was developed using ab259327, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling Dicer with ab259327 at 1/100 dilution (5.05 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebrum (PMID : 32737764). The section was incubated with ab259327 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

Lab

Western blot - Anti-Dicer antibody [EPR24104-105] - BSA and Azide free (AB280957)

This data was developed using ab259327, the same antibody clone in a different buffer formulation.

Lanes 1-4 : Merged signal (red and green). Green - ab259327 observed at 250 kDa. Red - Loading Control ab7291 observed at 50 kDa.

ab259327 was shown to specifically react with Dicer in wild-type HAP1 cells. Loss of signal was observed when Dicer knockout samples were used. Wild-type and Dicer knockout samples were subjected to SDS-PAGE. ab259327 and anti-alpha Tubulin - Loading Control ( ab7291) were incubated at 4℃ overnight 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-Dicer antibody [EPR24104-105] (<a href='/en-us/products/primary-antibodies/dicer-antibody-epr24104-105-ab259327'>ab259327</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

Dicer knockout HAP1 whole cell lysate at 20 µg

Lane 3:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 4:

2.4G2 (rat B cell lymphoma B lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 218 kDa

Observed band size: 250 kDa

false

• WB

Lab

Western blot - Anti-Dicer antibody [EPR24104-105] - BSA and Azide free (AB280957)

This data was developed using ab259327, the same antibody clone in a different buffer formulation.  False colour image of Western blot : Anti-Dicer antibody [EPR24104-105] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab259327 was shown to bind specifically to Dicer. A band was observed at 218 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in DICER1 knockout cell line. To generate this image, wild-type and DICER1 knockout HEK-293T 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.

Lanes 1 - 7:

Western blot - Anti-Dicer antibody [EPR24104-105] (<a href='/en-us/products/primary-antibodies/dicer-antibody-epr24104-105-ab259327'>ab259327</a>) at 1/1000 dilution

Lanes 1 - 7:

Western blot - Anti-Dicer antibody [EPR24104-105] - BSA and Azide free (ab280957) at 1/1000 dilution

Lane 1:

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

Lane 2:

DICER1 knockout HEK-293T cell lysate at 20 µg

Lane 3:

DICER1 4-25 knockout HEK-293T cell lysate at 20 µg

Lane 4:

Empty

Lane 5:

HeLa cell lysate at 20 µg

Lane 6:

Wild-type HAP1 cell lysate at 20 µg

Lane 7:

Dicer1 knockout HAP1 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

Observed band size: 218 kDa

false

• WB

Lab

Western blot - Anti-Dicer antibody [EPR24104-105] - BSA and Azide free (AB280957)

This data was developed using ab259327, the same antibody clone in a different buffer formulation.  False colour image of Western blot : Anti-Dicer antibody [EPR24104-105] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab259327 was shown to bind specifically to Dicer. A band was observed at 218 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in DICER1 knockout cell line. To generate this image, wild-type and DICER1 knockout HEK-293T 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.

Lanes 1 - 7:

Western blot - Anti-Dicer antibody [EPR24104-105] (<a href='/en-us/products/primary-antibodies/dicer-antibody-epr24104-105-ab259327'>ab259327</a>) at 1/1000 dilution

Lanes 1 - 7:

Western blot - Anti-Dicer antibody [EPR24104-105] - BSA and Azide free (ab280957) at 1/1000 dilution

Lane 1:

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

Lane 2:

DICER1 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human DICER1 knockout HEK293T cell line[2-20] (<a href='/en-us/products/cell-lines/human-dicer1-knockout-hek293t-cell-line2-20-ab291116'>ab291116</a>)

Lane 3:

DICER1 2-20 knockout HEK-293T cell lysate at 20 µg

Lane 4:

Empty

Lane 5:

HeLa cell lysate at 20 µg

Lane 6:

Wild-type HAP1 cell lysate at 20 µg

Lane 7:

Dicer1 knockout HAP1 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

Observed band size: 218 kDa

false

• WB

Supplier Data

Western blot - Anti-Dicer antibody [EPR24104-105] - BSA and Azide free (AB280957)

This data was developed using ab259327, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Lysates were made freshly and used in WB test immediately to minimize protein degradation.

Exposure time : Lane 1, 3, 4 : 3 minutes

Lane 2 : 26 seconds

All lanes:

Western blot - Anti-Dicer antibody [EPR24104-105] (<a href='/en-us/products/primary-antibodies/dicer-antibody-epr24104-105-ab259327'>ab259327</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 218 kDa

Observed band size: 250 kDa

false

• WB

Lab

Western blot - Anti-Dicer antibody [EPR24104-105] - BSA and Azide free (AB280957)

This data was developed using ab259327, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Negative control : THP-1 (PMID : 20584909).

Lysates were made freshly and used in WB test immediately to minimize protein degradation.

Exposure time : 48 seconds

All lanes:

Western blot - Anti-Dicer antibody [EPR24104-105] (<a href='/en-us/products/primary-antibodies/dicer-antibody-epr24104-105-ab259327'>ab259327</a>) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

2.4G2 (rat B cell lymphoma B lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 218 kDa

Observed band size: 250 kDa

false