Anti-NuMA 抗体 [EP3976]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NuMA antibody [EP3976] (AB109262)

ab109262, at 1/250 dilution, staining NuMA in paraffin-embedded Human breast tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NuMA antibody [EP3976] (AB109262)

ab109262, at 1/250 dilution, staining NuMA in paraffin-embedded Human colonic adenocarcinoma by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-NuMA antibody [EP3976] (AB109262)

ab109262, at 1/100 dilution, staining NuMA in HeLa cells by Immunofluorescence.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NuMA antibody [EP3976] (AB109262)

ab109262, at 1/250 dilution, staining NuMA in paraffin-embedded Human breast carcinoma by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NuMA antibody [EP3976] (AB109262)

Immunohistochemical analysis of paraffin-embedded mouse pancreatic cancer tissue labeling NuMA with ab109262 at 1/5000 dilution.

Nuclear staining on mouse pancreatic cancer.

The section was incubated with ab109262 for 10 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NuMA antibody [EP3976] (AB109262)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling NuMA with ab109262 at 1/5000 dilution.

Nuclear staining on mouse testis.

The section was incubated with ab109262 for 10 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NuMA antibody [EP3976] (AB109262)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling NuMA with ab109262 at 1/5000 dilution.

Nuclear staining on mouse kidney.

The section was incubated with ab109262 for 10 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-NuMA antibody [EP3976] (AB109262)

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) labelling NuMA with ab109262 at 1/500 (4.12 μg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody as secondary at 1/1000 (2 μg/ml) dilution. Confocal image showing nuclear and weak cytoplasmic staining in NIH/3T3 cell line (shown in green). ab7291 Anti-alpha Tubulin mouse monoclonal antibody at 1/1000 (1μg/ml) was used as a counterstain along with ab150120 as secondary at 1/1000 (2μg/ml) (shown in magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• WB

Unknown

Western blot - Anti-NuMA antibody [EP3976] (AB109262)

All lanes:

Western blot - Anti-NuMA antibody [EP3976] (ab109262) at 1/1000 dilution

Lane 1:

K562 cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

MCF7 cell lysate at 10 µg

Lane 4:

Raji cell lysate at 10 µg

Lane 5:

SW480 cell lysate at 10 µg

Lane 6:

CACO2 cell lysate at 10 µg

Lane 7:

MOLT4 cell lysate at 10 µg

Predicted band size: 238 kDa

false

• WB

Lab

Western blot - Anti-NuMA antibody [EP3976] (AB109262)

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

In Western blot, Anti-Vinculin antibody [EPR8185] - Loading Control (ab129002) staining at 1/5000 dilution (124KDa).

All lanes:

Western blot - Anti-NuMA antibody [EP3976] (ab109262) at 1/5000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 10 µg

Lane 2:

NIH/3T3 transfected with siRNA specifically targeting NuMA 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 1/100000 dilution

Observed band size: 238 kDa,124 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-NuMA antibody [EP3976] (AB109262)

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

In Western blot, Anti-Vinculin antibody [EPR8185] - Loading Control (ab129002) staining at 1/5000 dilution (124KDa).

The identity of the lower MW bands at approximately 100 + 120 kDa (in lanes 1-4) are unknown.

All lanes:

Western blot - Anti-NuMA antibody [EP3976] (ab109262) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse testis tissue lysate at 20 µg

Lane 4:

Mouse spleen tissue 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 1/100000 dilution

Observed band size: 238 kDa,124 kDa

false

Exposure time: 37s

• WB

CiteAb

Western blot - Anti-NuMA antibody [EP3976] (AB109262)

NuMA western blot using anti-NuMA antibody [EP3976] ab109262. Publication image and figure legend from Guo, L., Mohd, K. S., et al., 2019, J Cell Sci, PubMed 31434716.

ab109262 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 ab109262 please see the product overview.

Phosphorylation at T9 and S62 attenuates the interaction of importin-α1 with importin-β and NLS-containing SAFs in mitosis. (A) HeLa cells were transiently transfected with GFP, GFP–importin-α1 WT, GFP-importin-α1 2A or GFP–importin-α1 2D and were arrested in mitosis (labeled M) with 100 ng/ml of nocodazole for 17 h. GFP–importin-α1 was immunoprecipitated (IP) from mitotic cells using anti-GFP antibody, followed by immunoblot for importin-β, KIFC1, TPX2, NuMA and GFP. Cell lysates used for the precipitations are shown in the left panels. (B) HeLa cells were transiently transfected with either GFP, GFP–importin-α1 WT, GFP–importin-α1 2A or GFP–importin-α1 2D and were collected as asynchronous (labeled AS) cells. GFP–importin-α1 was immunoprecipitated from asynchronous cells using the anti-GFP antibody, followed by immunoblot for importin-β, KIFC1, TPX2, NuMA and GFP. Cell lysates used for the precipitations are shown in the left panels. (C) T9 and S62 phosphorylation may function in an NLS-dependent manner in mitosis. HeLa cells were co-transfected with the NLS-containing fragment GFP–NT and Myc-tagged importin-α1 or mutants [Myc-importin-α1 WT (myc-WT), the non-phosphorylation-mimicking double-mutant Myc-importin-α1 2A (myc-2A) and phosphorylation-mimicking double-mutant Myc-importin-α1 2D (myc-2D)] and then were arrested in mitosis with 100 ng/ml of nocodazole for 17 h. GFP–NT was immunoprecipitated from mitotic cells using an anti-GFP antibody, followed by immunoblot for Myc and GFP. (D) HeLa cells were co-transfected with the NLS-containing fragment GFP–NT and Myc-tagged importin-α1 or mutants as in C, and then were collected as asynchronous cells. GFP–NT was immunoprecipitated from asynchronous cells using an anti-GFP antibody, followed by immunoblot for Myc and GFP. The numbers under the bands refer to the relative grey value intensity.

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