Anti-BCL2L15 antibody [EPR17873]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-BCL2L15 antibody [EPR17873] (AB184932)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell line transfected with BCL2L15 (GFP-Tag) labeling BCL2L15 with ab184932 at 1/300 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG (PE) at 1/150 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-BCL2L15 antibody [EPR17873] (AB184932)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HEK-293T (human embryonic kidney) cells untransfected or transfected with with BCL2L15 expression vector labeling BCL2L15 with ab184932 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic staining on HEK-293T cells transfected with BCL2L15 expression vector.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and AlexaFluor^® 594 Goat anti-Mouse (ab150120) at 1/500 dilution (red).

The negative controls are as follows;
-ve control 1 : ab184932 at 1/500 dilution followed by AlexaFluor^® 594 Goat anti-Mouse (ab150120) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/500 dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-BCL2L15 antibody [EPR17873] (AB184932)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HEK293T (human embryonic kidney) cells transfected with GFP-BCL2L15 expression construct labeling BCL2L15 with ab184932 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) (ab150080) secondary antibody at 1/500 dilution (red). Confocal image showing cytoplasmic and weak nuclear staining on HEK293T transfected with GFP-BCL2L15 expression construct.

The nuclear counter stain is DAPI (blue). GFP is detected with an anti-GFP antibody (green) at 1/500 dilution.

-ve control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) (ab150080) secondary antibody at 1/500 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCL2L15 antibody [EPR17873] (AB184932)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling BCL2L15 with ab184932 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on human colon epithelium is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCL2L15 antibody [EPR17873] (AB184932)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling BCL2L15 with ab184932 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative control : no staining on human liver.

Counter stained with hematoxylin.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCL2L15 antibody [EPR17873] (AB184932)

Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling BCL2L15 with ab184932 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Negative control : no staining on human endometrium.

Counter stained with hematoxylin.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BCL2L15 antibody [EPR17873] (AB184932)

Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma tissue labeling BCL2L15 with ab184932 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and weak cytoplasm staining on the human gastric adenocarcinoma is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

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Western blot - Anti-BCL2L15 antibody [EPR17873] (AB184932)

Blocking/Dilution buffer : 5% NFDM/TBST.

The observed molecular mass of 45 kDa includes the 18 kDa BCL2L15 peptide and the 27 kDa GFP tag.

All lanes:

Western blot - Anti-BCL2L15 antibody [EPR17873] (ab184932) at 1/5000 dilution

Lane 1:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) non-transfected whole cell lysate at 10 µg

Lane 2:

HEK-293T transfected with a GFP-tagged BCL2L15 expression vector whole cell lysate at 10 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 1/1000 dilution

Predicted band size: 17 kDa

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Exposure time: 15s