Anti-SP140 antibody [EPR25120-83]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SP140 antibody [EPR25120-83] (AB314211)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HuT-78 (human Sezary syndrome cutaneous T lymphocyte) cells labelling SP140 with ab314211 at 1/500 dilution (0.1 ug)/Red (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, ab150081) at 1/5000 dilution was used as the secondary antibody.

• WB

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Western blot - Anti-SP140 antibody [EPR25120-83] (AB314211)

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

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-SP140 antibody [EPR25120-83] (ab314211) at 1/1000 dilution

Lane 1:

HuT-78 (human Sezary syndrome cutaneous T lymphocyte) whole cell lysate at 20 µg

Lane 2:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

Ramos (human Burkitts 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

Observed band size: 140 kDa,130 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-SP140 antibody [EPR25120-83] (AB314211)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 30102396, 28783698, 12368356).

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

All lanes:

Western blot - Anti-SP140 antibody [EPR25120-83] (ab314211) at 1/1000 dilution

Lane 1:

HuT-78 (human Sezary syndrome cutaneous T lymphocyte) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HuT-78 transfected with siRNA specifically targeti SP140 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

Observed band size: 140 kDa,130 kDa

false

Exposure time: 180s

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SP140 antibody [EPR25120-83] (AB314211)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling SP140 with ab314211 at 1/100 (5.0 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human tonsil.The section was incubated with ab314211 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). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SP140 antibody [EPR25120-83] (AB314211)

Immunohistochemical analysis of paraffin-embedded Human non-hodgkin's tissue labeling SP140 with ab314211 at 1/100 (5.0 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human non-hodgkin's lymphoma.The section was incubated with ab314211 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). 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-SP140 antibody [EPR25120-83] (AB314211)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling SP140 with ab314211 at 1/100 (5.0 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human kidney.The section was incubated with ab314211 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). 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-SP140 antibody [EPR25120-83] (AB314211)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling SP140 with ab314211 at 1/100 (5.0 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining on human cerebrum.The section was incubated with ab314211 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). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins