Anti-FUBP1/FBP antibody [EPR19208]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-FUBP1/FBP antibody [EPR19208] (AB213525)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling FUBP1/FBP with ab213525 at 1/600 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FUBP1/FBP antibody [EPR19208] (AB213525)

Immunohistochemical analysis of paraffin-embedded human oligodendroglioma tissue labeling FUBP1/FBP with ab213525 at 1/16000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on the vascular endothelium of human oligodendroglioma 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).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FUBP1/FBP antibody [EPR19208] (AB213525)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling FUBP1/FBP with ab213525 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FUBP1/FBP antibody [EPR19208] (AB213525)

Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling FUBP1/FBP with ab213525 at 1/16000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on the neurons of human cerebral cortex 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).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

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Immunoprecipitation - Anti-FUBP1/FBP antibody [EPR19208] (AB213525)

FUBP1/FBP was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab213525 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab213525 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Lane 1 : HeLa whole cell lysate 10μg (Input).

Lane 2 : ab213525 IP in HeLa whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab213525 in HeLa whole cell lysate.

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

Exposure time : 1 second.

The expression pattern observed is consistent with the literature (PMID : 24963357).

All lanes:

Immunoprecipitation - Anti-FUBP1/FBP antibody [EPR19208] (ab213525)

Predicted band size: 68 kDa

false

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FUBP1/FBP antibody [EPR19208] (AB213525)

Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling FUBP1/FBP with ab213525 at 1/16000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on the neurons of mouse cerebral cortex 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).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FUBP1/FBP antibody [EPR19208] (AB213525)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling FUBP1/FBP with ab213525 at 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FUBP1/FBP antibody [EPR19208] (AB213525)

Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling FUBP1/FBP with ab213525 at 1/16000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining on the neurons of rat stomach 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).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

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Western blot - Anti-FUBP1/FBP antibody [EPR19208] (AB213525)

Blocking/Dilution buffer : 5% NFDM/TBST.

The expression pattern observed is consistent with the literature (PMID : 24963357).

All lanes:

Western blot - Anti-FUBP1/FBP antibody [EPR19208] (ab213525) at 1/5000 dilution

Lane 1:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 10 µg

Lane 2:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 3:

Raji (Human Burkitt's lymphoma cell line) 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 1/100000 dilution

Predicted band size: 68 kDa

Observed band size: 74 kDa

false

Exposure time: 1s

• WB

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Western blot - Anti-FUBP1/FBP antibody [EPR19208] (AB213525)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1-3/5/6 : 1 second; Lane 4 : 3 seconds.

The expression pattern observed is consistent with the literature (PMID : 24963357).

Lanes 1 - 2:

Western blot - Anti-FUBP1/FBP antibody [EPR19208] (ab213525) at 1/1000 dilution

Lanes 3 - 6:

Western blot - Anti-FUBP1/FBP antibody [EPR19208] (ab213525) at 1/5000 dilution

Lane 1:

Rat brain lysate at 10 µg

Lane 2:

Mouse brain lysate at 10 µg

Lane 3:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 4:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 5:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

Lane 6:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) 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 1/100000 dilution

Predicted band size: 68 kDa

Observed band size: 74 kDa

false