Anti-HUWE1/Mule antibody [EPR24361-13]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HUWE1/Mule antibody [EPR24361-13] (AB271032)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labelling HUWE1/Mule with ab271032 at 1/1000 (0.525 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in human breast carcinoma (PMID : 29375730). The section was incubated with ab271032 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-HUWE1/Mule antibody [EPR24361-13] (AB271032)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling HUWE1/Mule with ab271032 at 1/50 (10.5 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green) Confocal image showing cytoplasmic and nuclear staining in HeLa cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-HUWE1/Mule antibody [EPR24361-13] (AB271032)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MOLT-4 (human lymphoblastic leukemia T lymphoblast, Left) and HeLa (Human cervix adenocarcinoma epithelial cell, Right) cells labelling HUWE1/Mule with ab271032 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Low expression : MOLT-4 (PMID : 15567145).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HUWE1/Mule antibody [EPR24361-13] (AB271032)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MOLT-4 cells labelling HUWE1/Mule with ab271032 at 1/50 (10.5 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing weak staining in MOLT-4 cells.Low expression : MOLT-4 (PMID : 15567145) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-HUWE1/Mule antibody [EPR24361-13] (AB271032)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized F9 (Mouse embryonal carcinoma epithelial cell) cells labelling HUWE1/Mule with ab271032 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HUWE1/Mule antibody [EPR24361-13] (AB271032)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labelling HUWE1/Mule with ab271032 at 1/1000 (0.525 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in rat kidney. The section was incubated with ab271032 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-HUWE1/Mule antibody [EPR24361-13] (AB271032)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized F9 cells labelling HUWE1/Mule with ab271032 at 1/50 (10.5 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). Confocal image showing cytoplasmic and nuclear staining in F9 cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HUWE1/Mule antibody [EPR24361-13] (AB271032)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labelling HUWE1/Mule with ab271032 at 1/1000 (0.525 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining in mouse cardiac muscle. The section was incubated with ab271032 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 LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

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

• WB

Lab

Western blot - Anti-HUWE1/Mule antibody [EPR24361-13] (AB271032)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The expression pattern & observed MW are consistent with what has been described in the literature (PMID : 25883150, 28938460).

Low expression : MOLT-4 (PMID : 15567145).

Exposure time : 5.5 seconds

All lanes:

Western blot - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032) at 1/1000 dilution

Lane 1:

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

Lane 2:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) 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 1/50000 dilution

Predicted band size: 481 kDa

Observed band size: 482 kDa

false

• WB

Lab

Western blot - Anti-HUWE1/Mule antibody [EPR24361-13] (AB271032)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTLysates were made freshly and used in WB test immediately to minimize protein degradation.

The expression pattern & observed MW are consistent with what has been described in the literature (PMID : 28938460).

Exposure time : Lane1 : 3.25 seconds. Lane2-3 : 5.5 seconds

All lanes:

Western blot - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

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 1/50000 dilution

Predicted band size: 481 kDa

Observed band size: 482 kDa

false

• WB

Lab

Western blot - Anti-HUWE1/Mule antibody [EPR24361-13] (AB271032)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The expression pattern & observed MW are consistent with what has been described in the literature (PMID : 28938460).

Exposure time : 10 seconds

All lanes:

Western blot - Anti-HUWE1/Mule antibody [EPR24361-13] (ab271032) at 1/1000 dilution

All lanes:

Mouse heart tissue 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 1/50000 dilution

Predicted band size: 481 kDa

Observed band size: 482 kDa

false