Anti-KLF4 抗体 [EPR19590]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-KLF4 antibody [EPR19590] (AB215036)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NCCIT (Human pluripotent embryonic carcinoma cell line) cells labeling KLF4 with ab215036 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on NCCIT 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.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody [EPR19590] (AB215036)

Immunohistochemical analysis of paraffin-embedded human seminoma tissue labeling KLF4 with ab215036 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Nuclear staining on tumor cells of human seminoma is observed [PMID : 17614846].

Counter stained with Hematoxylin.

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

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody [EPR19590] (AB215036)

Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling KLF4 with ab215036 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Nuclear staining on tumor cells of human gastric cancer is observed [PMID : 17614846].

Counter stained with Hematoxylin.

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

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody [EPR19590] (AB215036)

Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling KLF4 with ab215036 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Nuclear staining on human 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) at 1/500 dilution.

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

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-KLF4 antibody [EPR19590] (AB215036)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NCCIT (Human pluripotent embryonic carcinoma cell line) cells labeling KLF4 with ab215036 at 1/50 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.

• ChIP-seq

Lab

ChIP-sequencing - Anti-KLF4 antibody [EPR19590] (AB215036)

Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab215036 [EPR19590]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The input control is also shown.

• IP

Unknown

Immunoprecipitation - Anti-KLF4 antibody [EPR19590] (AB215036)

ab215036 (purified) at 1/30 dilution immunoprecipitating KLF4 in NCCIT whole cell lysate.
Lane 1 (input) : NCCIT (Human pluripotent embryonic carcinoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+) : ab215036 & NCCIT whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab215036 in NCCIT whole cell lysate
For western blotting, ab215036 at 1/500 dilution (0.117 µg/ml) and veriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution were used.
Blocking and diluting buffer : 5% NFDM /TBST .

All lanes:

Immunoprecipitation - Anti-KLF4 antibody [EPR19590] (ab215036)

Predicted band size: 15 kDa,54 kDa

false

• ChIP-seq

Lab

ChIP-sequencing - Anti-KLF4 antibody [EPR19590] (AB215036)

Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab215036 [EPR19590]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-KLF4 antibody [EPR19590] (AB215036)

Chromatin was prepared from NCCIT cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab215036 [EPR19590]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The input control is also shown.

• WB

Supplier Data

Western blot - Anti-KLF4 antibody [EPR19590] (AB215036)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-KLF4 antibody [EPR19590] (ab215036) at 1/1000 dilution

All lanes:

NCCIT (Human pluripotent embryonic carcinoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 54 kDa

Observed band size: 54 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-KLF4 antibody [EPR19590] (AB215036)

Blocking buffer and concentration : 5% NFDM/TBST

Diluting buffer and concentration : 5% NFDM/TBST

Gel type : 4-20% gradient gel(SDS-PAGE)

All lanes:

Western blot - Anti-KLF4 antibody [EPR19590] (ab215036) at 1/1000 dilution

Lane 1:

GST tagged Recombinant Human KLF4 protein (Full length, 81 KDa)

Lane 2:

GST tagged Recombinant Human Adipose Triglyceride Lipase (PNPLA2) protein (Full length, 82 KDa)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 54 kDa

Observed band size: 81 kDa

false

Exposure time: 1s