PHF8 Antibody

Novus Biologicals | Catalog # NB100-93314

Novus Biologicals
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Key Product Details

Validated by

Knockout/Knockdown, Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Immunoprecipitation, Knockdown Validated

Cited:

Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
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Product Specifications

Immunogen

The immunogen recognized by this antibody maps to a region between residue 925 and 975 of human PHD finger protein 8 using the numbering given in entry CAI41576.1 (GeneID 23133).

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

Novus Biologicals Rabbit PHF8 Antibody (NB100-93314) is a polyclonal antibody validated for use in WB and IP. Anti-PHF8 Antibody: Cited in 2 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for PHF8 Antibody

Western Blot: PHF8 Antibody [NB100-93314]

Western Blot: PHF8 Antibody [NB100-93314]

Western Blot: PHF8 Antibody [NB100-93314] - Detection of Human PHF8 by Western Blot. Samples: Whole cell lysate (50 ug) from HeLa, HeLa (RIPA), 293T, and Jurkat cells. Antibodies: Affinity purified rabbit anti-PHF8 antibody NB100-93314 used for WB at 0.04 ug/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Knockdown Validated: PHF8 Antibody [NB100-93314]

Western Blot: PHF8 Antibody [NB100-93314]

PHF8-Antibody-Western-Blot-NB100-93314-img0009.jpg
PHF8 Antibody

Western Blot: PHF8 Antibody [NB100-93314] -

Western Blot: PHF8 Antibody [NB100-93314] - PHF8 interacts with the AR & functions as an AR coactivator. (a) PHF8 interacts with AR independent of DHT. HEK293T cells were transiently transfected with plasmids expressing Flag-tagged PHF8 & GFP-tagged AR. Cells were treated with or without 10 nM DHT for 48 h & the cell lysates were used for immunoprecipitation with antibodies against either Flag or GFP. Western blotting was conducted with antibodies as indicated. (b) PHF8 interacts with AR independent of its demethylase activity. HEK293T cells were transiently transfected with plasmids expressing Flag-tagged PHF8 (H247A) mutant & GFP-tagged AR. The interaction assay by co-immunoprecipitation was performed as described above. (c) In LNCaP cells endogenous PHF8 interacts with AR. LNCaP cells were treated with 10 nM DHT for 48 h & the cell lysates were used for immunoprecipitation with antibodies against either PHF8 or AR. Western blotting was conducted with AR or PHF8 antibody as indicated. (d) PHF8 enhances AR transcriptional activation in a demethylase activity-dependent manner. HeLa cells were co-transfected with the 4 × UAS-TK-luc reporter, the plasmids expressing the fusion protein Gal-AR, & increased amount of either the wild-type PHF8 or mutant PHF8 (H247A) as indicated. Cells were cultured with or without 10 nM R1881 overnight. The relative luciferase activities in the cell lysates were normalized to renilla luciferase activity. The relative luciferase activities are presented as mean±s.e.m. of three independent transfections & the levels of PHF8 were estimated by western blot as depicted in the bottom panel. Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/oncsis201674), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
PHF8 Antibody

Knockdown Validated: PHF8 Antibody [NB100-93314] -

PHF8 is recruited to AR target gene promoters and is essential for optimal hormone-dependent transcriptional activation of AR target genes. (a, b) PHF8 is recruited to the AR target genes PSA (a) and NKX3.1 (b) upon DHT treatment in LNCaP cells. LNCaP cells were cultured in absence or presence of DHT for 48 h. Whole-cell lysates were subjected to ChIP assays with nonspecific IgG or antibodies against AR, PHF8 or RNA polymerase II (RNA-Pol). Purified DNA was used for qPCR with primers targeting the promoter regions of the PSA (Left Panel) and NKX3.1 (Right Panel). The mean values+/-s.e.m. were shown. Student's t-tests were performed to compare the assays with specific antibodies against AR, PHF8 and RNA-Pol, or nonspecific IgG. *P<0.05, compared with DHT-negative group; and #P<0.05, compared with IgG group. (c) Western blot analysis showing knockdown of PHF8 by shRNA against PHF8. LNCaP cells were infected with a lentiviral shPHF8 and western blot analysis was carried out with an anti-PHF8 antibody. (d) Knockdown PHF8 severely impaired DHT-induced transcriptional activation of PSA and NKX3.1. The control shRNA and shPHF8 infected LNCaP cells were treated with 10 nM DHT for 24 h and total RNAs were prepared and subjected to quantitative RT–PCR analysis. The results are presented as the mean+/-s.e.m. of three independent experiments. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/27991916), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for PHF8 Antibody

Application
Recommended Usage

Western Blot

1:2000-1:10000
Application Notes
Use in immunoprecipitation reported in scientific literature (PMID: 27991916). Western blot of lysates performed using standard western blot reagents and 4-8% SDS-PAGE.

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

TBS and 0.1% BSA

Preservative

0.09% Sodium Azide

Concentration

0.2 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C. Do not freeze.

Background: PHF8

PHF8 (PHD finger protein 8) is a member of the jumonji family of proteins and contains a jumonji C (JmjC) domain. The JmjC proteins are predicted to be metalloenzymes that play a role a chromatin remodeling and histone demethylation. PHF8 also bears a PHD (plant homeodomain)- type zinc-finger, a domain also found to be involved in chromatin remodeling and transcriptional regulation. Truncating mutations in PHF8 are associated with X-linked mental retardation and cleft lip/cleft palate. PHF8 is also known as ZNF422 and KIAA1111

Long Name

Histone lysine demethylase PHF8

Alternate Names

KIAA1111, ZNF422

Entrez Gene IDs

23133 (Human)

Gene Symbol

PHF8

UniProt

Additional PHF8 Products

Product Documents for PHF8 Antibody

Certificate of Analysis

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Product Specific Notices for PHF8 Antibody

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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