Anti-HMGN1 antibody [EPR27314-67]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGN1 antibody [EPR27314-67] (AB315889)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling HMGN1 with ab315889 at 1/8000 (0.065 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen. The section was incubated with ab315889 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed 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-HMGN1 antibody [EPR27314-67] (AB315889)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling HMGN1 with ab315889 at 1/8000 (0.065 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human breast carcinoma. The section was incubated with ab315889 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed 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-HMGN1 antibody [EPR27314-67] (AB315889)

Immunohistochemical analysis of paraffin-embedded Human epityphlon tissue labeling HMGN1 with ab315889 at 1/8000 (0.065 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human epityphlon. The section was incubated with ab315889 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-HMGN1 antibody [EPR27314-67] (AB315889)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling HMGN1 with ab315889 at 1/500 (1.032 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 nuclear staining in HeLa cell line.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/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-HMGN1 antibody [EPR27314-67] (AB315889)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling HMGN1 with ab315889 at 1/500 dilution (0.1 ug)/(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).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• WB

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Western blot - Anti-HMGN1 antibody [EPR27314-67] (AB315889)

The identity of the bands above 50 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-HMGN1 antibody [EPR27314-67] (ab315889) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 40 µg with 5% NFDM/TBST

Lane 2:

HeLa transfected with siRNA specifically targeting HMGN1 whole cell lysate at 40 µg with 5% NFDM/TBST

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 1/20000 dilution

Observed band size: 17 kDa,36 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-HMGN1 antibody [EPR27314-67] (AB315889)

The identity of the bands between 50 kDa and 150 kDa are unknown.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-HMGN1 antibody [EPR27314-67] (ab315889) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 3:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

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 1/20000 dilution

Observed band size: 17 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-HMGN1 antibody [EPR27314-67] (AB315889)

The identity of the bands above 50 kDa are unknown.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-2 : 15 seconds, lane 3 : 1 second.

All lanes:

Western blot - Anti-HMGN1 antibody [EPR27314-67] (ab315889) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 3:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

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 1/20000 dilution

Observed band size: 17 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-HMGN1 antibody [EPR27314-67] (AB315889)

This antibody does not cross-react with human HMGN2, HMGN3, HMGN4, HMGN5.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-HMGN1 antibody [EPR27314-67] (ab315889) at 1/1000 dilution

Lane 1:

His-tag human HMGN1 recombinant protein at 100 ng with 5% NFDM/TBST

Lane 2:

His-tag human HMGN2 recombinant protein at 100 ng with 5% NFDM/TBST

Lane 3:

His-tag human HMGN3 recombinant protein at 100 ng with 5% NFDM/TBST

Lane 4:

His-tag human HMGN4 recombinant protein at 100 ng with 5% NFDM/TBST

Lane 5:

His-tag human HMGN5 recombinant protein at 100 ng with 5% NFDM/TBST

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

Observed band size: 25 kDa

true

Exposure time: 180s