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AB261722

Human PHF6 (PHD finger protein 6) knockout HEK-293 cell line

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PHF6 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp deletion.

View Alternative Names

AC004383.6, BFLS, BORJ, CENP 31, Centromere protein 31, MGC14797, OTTHUMP00000024063, PHD finger protein 6, PHD-like zinc finger protein, PHF6_HUMAN

3 Images
Western blot - Human PHF6 (PHD finger protein 6) knockout HEK-293 cell line (AB261722)
  • WB

Lab

Western blot - Human PHF6 (PHD finger protein 6) knockout HEK-293 cell line (AB261722)

Lanes 1 - 4 : Merged signal (red and green). Green - ab173304 observed at 41 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab173304 was shown to recognize PHD finger protein 6 in wild-type HEK-293 cells as signal was lost at the expected MW in PHF6 knockout cell line ab261722 (knockout cell lysate ab261661). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and PHF6 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab173304 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PHD finger protein 6/PHF6 antibody [EPR11997] (<a href='/en-us/products/primary-antibodies/phd-finger-protein-6-phf6-antibody-epr11997-ab173304'>ab173304</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

PHF6 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 41 kDa

false

Western blot - Human PHF6 (PHD finger protein 6) knockout HEK-293 cell line (AB261722)
  • WB

Lab

Western blot - Human PHF6 (PHD finger protein 6) knockout HEK-293 cell line (AB261722)

Lanes 1 - 4 : Merged signal (red and green). Green - ab170929 observed at 41 kDa. Red - loading control ab130007 observed at 130 kDa.

ab170929 was shown to recognize PHD finger protein 6/PHF6 in wild-type HEK 293 cells as signal was lost at the expected MW in PHF6 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PHF6 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. ab170929 and ab130007 (Mouse anti Vinculin loading control) were incubated overnight at 4° at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PHD finger protein 6/PHF6 antibody [EPR11996(B)] (<a href='/en-us/products/primary-antibodies/phd-finger-protein-6-phf6-antibody-epr11996b-ab170929'>ab170929</a>) at 1/5000 dilution

Lane 1:

Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

PHF6 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

Western blot - Human PHF6 (PHD finger protein 6) knockout HEK-293 cell line (ab261722)

Lane 3:

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

Lane 4:

A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 41 kDa

false

Next Generation Sequencing - Human PHF6 (PHD finger protein 6) knockout HEK-293 cell line (AB261722)
  • NGS

Lab

Next Generation Sequencing - Human PHF6 (PHD finger protein 6) knockout HEK-293 cell line (AB261722)

1 bp deletion after Arg15 of the WT protein

Key facts

Cell type

HEK-293

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 1 bp deletion

Reactivity data

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Product details

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

What's included?

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Properties and storage information

Gene name
PHF6
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

DMEM (High Glucose) + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

PHD finger protein 6 also known as PHF6 is a protein with a molecular mass of approximately 43 kDa. It consists of two PHD domains which are zinc finger-like motifs playing roles in chromatin remodeling. PHF6 expresses widely in various tissues with noticeable levels in the brain and hematopoietic cells indicating its significance in these areas. Alternate names for PHF6 include ZNF312 and BORJ although PHF6 is the most commonly referred designation.
Biological function summary

PHF6 involves a range of cellular processes including regulation of transcription and chromatin structure modification. PHF6 can bind to certain DNA sequences and proteins to influence gene expression. It interacts with chromatin remodeling complexes indicating a role in maintaining chromatin accessibility for transcription factors. The protein's PHD domains contribute to its ability to engage in these interactions impacting the transcriptional activity of target genes.

Pathways

PHF6 interacts with transcriptional and chromatin remodeling pathways. It plays a role in the interplay between chromatin and transcriptional machinery which is critical for regulating gene expression. PHF6 associates with proteins such as nucleophosmin and other chromatin regulators influencing pathways like the Notch signaling pathway which is vital for cellular differentiation and development. These interactions highlight its involvement in genetic regulation within developmental pathways.

Mutations in PHF6 link to Börjeson-Forssman-Lehmann syndrome (BFLS) a rare genetic disorder affecting neurological development. Additionally PHF6 mutations recurrently appear in T-cell acute lymphoblastic leukemia (T-ALL) highlighting a role in hematological malignancies. In the context of T-ALL PHF6 interacts with proteins like Notch1 which drives leukemic proliferation when dysregulated. These associations make PHF6 a significant focus for understanding and potentially targeting specific genetic disorders and cancers.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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