Anti-FGF2 antibody [EPR20145-219]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FGF2 antibody [EPR20145-219] (AB208687)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized SK-OV-3 (Human ovarian cancer cell line) cells labeling FGF2 with ab208687 at 1/500 dilution followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear weak cytoplasmic staining on SK-OV-3 cell line.

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

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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-FGF2 antibody [EPR20145-219] (AB208687)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed K562 (Human chronic myelogenous leukemia cell line from bone marrow) cells labeling FGF2 with ab208687 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FGF2 antibody [EPR20145-219] (AB208687)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling FGF2 with ab208687 at 1/500 dilution followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear and weak cytoplasmic staining on U-87 MG cell line.

The nuclear counterstain is DAPI (blue).

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

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

• IP

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Immunoprecipitation - Anti-FGF2 antibody [EPR20145-219] (AB208687)

FGF2 was immunoprecipitated from 0.35 mg of K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate with ab208687 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab208687 at 1/500 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : K562 whole cell lysate 10μg (Input).

Lane 2 : ab208687 IP in K562 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab208687 in K562 whole cell lysate.

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

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-FGF2 antibody [EPR20145-219] (ab208687)

Predicted band size: 30 kDa

false

• I-ELISA

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Indirect ELISA - Anti-FGF2 antibody [EPR20145-219] (AB208687)

ELISA analysis of Mouse FGF2 recombinant protein at 1000 ng/ml with ab208687. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

• WB

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Western blot - Anti-FGF2 antibody [EPR20145-219] (AB208687)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 : 3 minutes; Lane 2 : 15 seconds; Lane 3 : 5 seconds.

The MW observed is consistent with the literatures (PMID : 18850066, PMID : 24827991).

All lanes:

Western blot - Anti-FGF2 antibody [EPR20145-219] (ab208687) at 1/1000 dilution

Lane 1:

K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 10 µg

Lane 2:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 10 µg

Lane 3:

SK-OV-3 (Human ovarian cancer 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: 30 kDa

Observed band size: 18 kDa,22 kDa,24 kDa

false

• WB

Supplier Data

Western blot - Anti-FGF2 antibody [EPR20145-219] (AB208687)

Blocking/Dilution buffer : 5% NFDM/TBST.

The MW observed is consistent with the literatures (PMID : 18850066, PMID : 24827991).

All lanes:

Western blot - Anti-FGF2 antibody [EPR20145-219] (ab208687) at 1/1000 dilution

Lane 1:

Human fetal kidney lysate at 10 µg

Lane 2:

Human prostate lysate at 10 µg

Lane 3:

Human fetal heart lysate at 10 µg

Lane 4:

Human testis lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 30 kDa

Observed band size: 18 kDa,22 kDa,24 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-FGF2 antibody [EPR20145-219] (AB208687)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-FGF2 antibody [EPR20145-219] (ab208687) at 1/10000 dilution

All lanes:

Recombinant human FGF2 active protein (aa143-288) at 0.01 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: 30 kDa

Observed band size: 15 kDa

false

Exposure time: 1s