Anti-PRPF4 antibody [EPR17206(B)]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PRPF4 antibody [EPR17206(B)] (AB201684)

Intracellular Flow Cytometry analysis of HeLa cells labelling PRPF4 (red) with purified ab201684 at dilution of 1/50. The secondary antibody used was Alexa Fluorr^® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PRPF4 antibody [EPR17206(B)] (AB201684)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PRPF4 with ab201684 at 1/1200 dilution followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

The negative controls are as follows :
-ve control 1 : ab201684 at 1/1200 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PRPF4 antibody [EPR17206(B)] (AB201684)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling PRPF4 with ab201684 at 1/1200 dilution followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on MCF7 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

The negative controls are as follows :
-ve control 1 : ab201684 at 1/1200 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRPF4 antibody [EPR17206(B)] (AB201684)

Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling PRPF4 with ab201684 at 1/3000 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on Human cervix carcinoma tissue 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.

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

• IP

Supplier Data

Immunoprecipitation - Anti-PRPF4 antibody [EPR17206(B)] (AB201684)

PRPF4 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab201684 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab201684 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1500 dilution.

Lane 1 : HeLa whole cell lysate 10 μg (Input). Lane 2 : ab201684 IP in HeLa whole cell lysate. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab201684 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 5 seconds.

All lanes:

Immunoprecipitation - Anti-PRPF4 antibody [EPR17206(B)] (ab201684)

Predicted band size: 58 kDa

false

• WB

Supplier Data

Western blot - Anti-PRPF4 antibody [EPR17206(B)] (AB201684)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PRPF4 antibody [EPR17206(B)] (ab201684) at 1/5000 dilution

Lane 1:

HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

SW480 (Human colon adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 3:

Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 4:

Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 58 kDa

Observed band size: 58 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-PRPF4 antibody [EPR17206(B)] (AB201684)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PRPF4 antibody [EPR17206(B)] (ab201684) at 1/5000 dilution

All lanes:

Human fetal spleen lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 58 kDa

Observed band size: 58 kDa

false

Exposure time: 3min