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AB285360

Human GATA1 knockout K562 cell line[C116]

Human GATA1 knockout K562 cell line[C116]

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GATA1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Sanger sequencing of this cell line reveals two indels with equal contribution: -34bp and -36bp from the beginning of exon 2, spanning across the intron/exon border (clone 67). Knockout. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
1 Images
Western blot - Human GATA1 knockout K562 cell line[C116] (AB285360)
  • WB

Lab

Western blot - Human GATA1 knockout K562 cell line[C116] (AB285360)

Western blot : Anti-GATA1 antibody [EPR17362] (ab181544) staining at 1/10000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab181544 was shown to bind specifically to GATA1. A band was observed at 45 kDa in wild-type K562 cell lysates with no signal observed at this size in GATA1 knockout cell line ab285360 (knockout K562 cell lysate). To generate this image, wild-type and GATA1 knockout K562 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-GATA1 antibody [EPR17362] - ChIP Grade (<a href='/products/primary-antibodies/gata1-antibody-epr17362-chip-grade-ab181544'>ab181544</a>) at 1/10000 dilution

Lane 1:

Wild-type K562 cell lysate at 20 µg

Lane 2:

GATA1 knockout K562 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 45 kDa

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Key facts

細胞タイプ

K-562

生物種

Human

組織

Blood

製品の状態

Liquid

form

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

Sanger sequencing of this cell line reveals two indels with equal contribution: -34bp and -36bp from the beginning of exon 2, spanning across the intron/exon border (clone 67). Knockout.

疾病

Chronic Myelogenous Leukemia

Reactivity data

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

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

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出荷温度及び保存条件

遺伝子名
GATA1
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
Sanger Sequencing, Western blot
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 for bath 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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 2x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. 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 2x105 cells/mL is recommended.
  • Subculture when cells reach 1x106 cells/mL.
培養培地

IMDM + 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.

GATA1 also known as GATA-binding factor 1 is a transcription factor weighing approximately 42 kDa. This protein binds to the DNA sequence "GATA" within the regulatory regions of target genes to control their expression. GATA1 is mostly expressed in hematopoietic cells especially erythroid cells megakaryocytes eosinophils and mast cells. Its specific expression pattern allows it to regulate genes essential for the development and function of these cell types.
Biological function summary

GATA1 regulates gene expression and is essential for erythroid and megakaryocyte differentiation. GATA1 belongs to a family of GATA transcription factors and can form regulatory complexes with other proteins such as FOG1 (Friend of GATA1) and TAL1. These complexes facilitate the binding of GATA1 to DNA enhancing its transcriptional activity on target gene sets necessary for blood cell maturation and function.

Pathways

GATA1 participates in the erythropoiesis and megakaryopoiesis pathways. It directly regulates several genes and coordinates signals with factors like GATA2 and RUNX1 which are other key proteins involved in these pathways. Through these interactions GATA1 ensures the correct development of red blood cells and platelets maintaining hematopoietic homeostasis in the organism.

Abnormal GATA1 function is linked to blood-related disorders. Mutations or dysregulations of GATA1 can lead to diseases such as X-linked thrombocytopenia with dyserythropoietic anemia which involves low platelet count and ineffective red blood cell production. Moreover GATA1 mutations are associated with certain leukemias particularly acute megakaryoblastic leukemia. GATA1's dysfunction in these diseases often occurs in conjunction with abnormalities in related proteins like GATA2 influencing disease progression and phenotype.

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Suspension

Gender

Female

製品プロトコール

ターゲットの情報

See full target information GATA1

文献 (1)

Recent publications for all applications. Explore the full list and refine your search

Molecular therapy. Nucleic acids 7:288-298 PubMed28624204

2017

CRISPR/Cas9-Directed Reassignment of the GATA1 Initiation Codon in K562 Cells to Recapitulate AML in Down Syndrome.

Applications

Unspecified application

Species

Unspecified reactive species

Kevin M Bloh,Pawel A Bialk,Anilkumar Gopalakrishnapillai,E Anders Kolb,Eric B Kmiec
View all publications

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