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AB286487

Human HDAC4 knockout HCT116 cell line

Human HDAC4 knockout HCT116 cell line

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HDAC4 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.

別名を表示する

AHO3, BDMR, EC 3.5.1.98, HA6116, HD 4, HDAC A, HDAC4_HUMAN, Histone Deacetylase A, Histone deacetylase 4, KIAA0288

2 Images
Western blot - Human HDAC4 knockout HCT116 cell line (AB286487)
  • WB

Lab

Western blot - Human HDAC4 knockout HCT116 cell line (AB286487)

Western blot : Rabbit Monoclonal [EPR22937-157] to HDAC4 ab235583 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 137 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in HDAC4 knockout HCT 116 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-HDAC4 antibody [EPR22937-157] (<a href='/products/primary-antibodies/hdac4-antibody-epr22937-157-ab235583'>ab235583</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 at 20 µg

Lane 2:

HDAC4 knockout HCT 116 at 20 µg

Lane 2:

Western blot - Human HDAC4 knockout HCT116 cell line (ab286487) at 20 µg

Lane 3:

Wild-type U-87 MG at 20 µg

Lane 4:

HDAC4 knockout U-87 MG at 20 µg

Lane 5:

Wild-type A549 at 20 µg

Lane 6:

HDAC4 knockout A549 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 119 kDa

Observed band size: 137 kDa

false

Next Generation Sequencing - Human HDAC4 knockout HCT116 cell line (AB286487)
  • NGS

Lab

Next Generation Sequencing - Human HDAC4 knockout HCT116 cell line (AB286487)

277 bp deletion after Glu337

Key facts

細胞タイプ

HCT116

生物種

Human

組織

Colon

製品の状態

Liquid

form

ノックアウト検証方法

Next Generation Sequencing

疾病

Carcinoma

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

製品の詳細

Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.

Recommended control: Human wild-type HCT116 cell line (ab288559). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

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.

製品内容

{ "values": { "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab286487-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab286487 Human HDAC4 knockout HCT116 cell line", "number":"AB286487-CMP01", "productcode":"" } ] } } }

出荷温度及び保存条件

遺伝子名
HDAC4
遺伝子編集のタイプ
Knockout
遺伝子編集の方法
CRISPR technology
ノックアウト検証方法
Next Generation Sequencing
出荷温度
Dry Ice
短期保存温度
-196°C
長期保存温度
-196°C

取り扱い方法

初回取り扱いガイドライン

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.

継代培養ガイドライン
  • 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.
培養培地

McCoY5a + 10% FBS

凍結保存培地

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

補足情報

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

HDAC4 also known as Histone Deacetylase 4 functions in the regulation of gene expression through chromatin remodeling. This protein removes acetyl groups from histone proteins causing chromatin compaction and transcriptional repression. HDAC4 has a molecular weight of approximately 140 kDa. The protein is found in various tissues including brain heart and skeletal muscle and plays essential roles in various cellular processes. Alternate names for HDAC4 include KIAA0288 and HD4.
Biological function summary

The role of HDAC4 extends beyond chromatin remodeling as it serves as a critical regulator in several cellular functions. As part of a complex HDAC4 interacts with other proteins to control cell cycle differentiation and apoptosis. It cooperates with transcriptional repressors like MEF2 and RUNX2 influencing various signaling pathways. The protein's activity impacts neuronal survival muscle development and cardiac hypertrophy through its regulatory mechanisms.

Pathways

HDAC4 integrates into significant signaling networks notably the MAPK and calcium-calmodulin signaling pathways. Within the MAPK pathway HDAC4 associates with proteins like MEF2 impacting cellular growth and differentiation processes. In the calcium-calmodulin pathway HDAC4 affects gene transcription via its interaction with the phosphatase calcineurin linking intracellular calcium levels with transcriptional responses.

HDAC4 has associations with neurological disorders and cancer. Mutations and dysregulation in HDAC4 are linked to neuronal disorders such as Huntington's disease. In cancer HDAC4 interacts with other oncogenic proteins such as p53 influencing tumor progression and resistance to therapy. Consequently HDAC4 presents as a potential target for therapeutic intervention in these disease contexts.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

製品プロトコール

Abcam product promise

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