Anti-PHD finger protein 1/PHF1 antibody [EPR14222]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (AB184951)

Intracellular flow cytometric analysis of 293 cells (2% paraformaldehyde-fixed) labeling PHF1with ab184951 at 1/160 dilution (red) mpared to a negative control antibody (red)or a rabbit IgG (negative) (green), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (AB184951)

Immunofluorescent analysis of MCF7 cells (4% paraformaldehyde-fixed) labeling PHF1 with ab184951 at 1/500 dilution followed by Goat anti rabbit IgG (AlexaFluor® 555) secondary at 1/200 dilution and counter-stained with DAPI (blue).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (AB184951)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling PHF1 with ab184951 at 1/100 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (AB184951)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling PHF1 with ab184951 at 1/100 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• IP

Lab

Immunoprecipitation - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (AB184951)

PHF1 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate 10 ug with ab184951 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab184951 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse brain tissue lysate 10 ug

Lane 2 : ab184951 IP in Mouse brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab184951 in mouse brain tissue lysate

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

Exposure time : 3 minutes

All lanes:

Immunoprecipitation - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (ab184951)

Predicted band size: 62 kDa

false

• WB

Supplier Data

Western blot - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (AB184951)

All lanes:

Western blot - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (ab184951) at 1/1000 dilution

Lane 1:

Mouse brain tissue at 20 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 3:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 4:

Rat brain tissue lysate at 20 µg

Lane 5:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 6:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 62 kDa

false

• WB

Supplier Data

Western blot - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (AB184951)

All lanes:

Western blot - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (ab184951) at 1/5000 dilution

All lanes:

Human fetal brain lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 62 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (AB184951)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ NCCIT (human pluripotent embryonic carcinoma epithelial cell) cells and 5 µg of ab184951 [EPR14222]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (AB184951)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ NCCIT (human pluripotent embryonic carcinoma epithelial cell) cells and 5 µg of ab184951 [EPR14222]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (AB184951)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10⁵ NCCIT (human pluripotent embryonic carcinoma epithelial cell) cells and 5 µg of ab184951 [EPR14222]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• WB

Supplier Data

Western blot - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (AB184951)

All lanes:

Western blot - Anti-PHD finger protein 1/PHF1 antibody [EPR14222] (ab184951) at 1/20000 dilution

Lane 1:

293 lysate at 10 µg

Lane 2:

Jurkat lysate at 10 µg

Lane 3:

MCF7 lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 62 kDa

false