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AB300930

Human ERRFI1 knockout A549 cell line

Human ERRFI1 knockout A549 cell line

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ERRFI1 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

別名を表示する

ERBB receptor feedback inhibitor 1, ERRFI_HUMAN, Errfi1, GENE 33, Gene 33, rat, homolog of, Mig-6, Mitogen-inducible gene 6 protein, RALT, mitogen inducible gene 6, receptor-associated late transducer

2 Images
Western blot - Human ERRFI1 knockout A549 cell line (AB300930)
  • WB

Lab

Western blot - Human ERRFI1 knockout A549 cell line (AB300930)

Western blot : Anti-ERRFI1 antibody (ab227944) staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab227944 was shown to bind specifically to ERRFI1. A band was observed at 51 kDa in wild-type A549 cell lysates with no signal observed at this size in ERRFI1 knockout cell line. To generate this image, wild-type and ERRFI1 knockout A549 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-MIG-6 antibody (<a href='/products/primary-antibodies/mig-6-antibody-ab227944'>ab227944</a>) at 1/2000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

ERRFI1 knockout A549 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: 51 kDa

false

Next Generation Sequencing - Human ERRFI1 knockout A549 cell line (AB300930)
  • NGS

Lab

Next Generation Sequencing - Human ERRFI1 knockout A549 cell line (AB300930)

82 bp deletion after Lys 71 of WT protein

Key facts

細胞タイプ

A549

生物種

Human

組織

Lung

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>" }, "WB": { "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
ERRFI1
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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
培養培地

F-12K + 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.

MIG-6 also known as Gene 33 or ERRFI1 is a protein that functions as an adaptor/scaffold. It has an approximate mass of 70 kilodaltons and is found predominantly in epithelial tissues. MIG-6 acts as a negative regulator of the epidermal growth factor receptor (EGFR) signaling pathway. This regulation role involves binding to the tyrosine kinase domain of the EGFR which leads to a decrease in its catalytic activity. Besides its presence in epithelial tissues MIG-6 is also expressed in a variety of other tissue types including the liver and pancreas.
Biological function summary

MIG-6 plays a significant role in the regulation of cell proliferation differentiation and apoptosis. It is a part of a larger signal transduction network which is essential for controlling cellular responses to external stimuli. By acting as a brake on EGFR signaling MIG-6 helps to balance cell activities to prevent excessive growth and proliferation. The protein's impact extends to influencing the behavior of other signaling molecules within the cell contributing to meditative roles in maintaining cellular harmony.

Pathways

MIG-6 is integral to the EGFR signaling pathway and is closely associated with the MAPK (mitogen-activated protein kinase) pathway. Its regulatory function affects downstream signaling events that involve proteins like RAF MEK and ERK which are all critical in cell growth and survival. By interacting with these pathways MIG-6 modulates signal transduction cascades which allows for controlled cellular functions and responses. All these connections make it an important node for intracellular communication and pathway intersections.

Deficiencies or dysregulation of MIG-6 have been linked to cancer particularly in tumors that have heightened EGFR activity. The protein's ability to suppress EGFR signaling is essential in preventing unchecked cell proliferation that can lead to tumor growth. Additionally research has connected altered levels of MIG-6 with inflammatory disorders. Abnormal interactions between MIG-6 and EGFR can exacerbate inflammatory responses contributing to various chronic inflammatory conditions. Understanding these associations helps in the development of targeted therapies for conditions with altered EGFR signaling.

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

製品プロトコール

ターゲットの情報

See full target information ERRFI1

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