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AB300883

人CDA knockout A549 cell line

Human CDA knockout A549 cell line

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CDA KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. 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'.

查看别名

Cytidine aminohydrolase, Cytidine deaminase, EC 3.5.4.5

1 Images
Western blot - Human CDA knockout A549 cell line (AB300883)
  • WB

Lab

Western blot - Human CDA knockout A549 cell line (AB300883)

Western blot : Rabbit Monoclonal [EPR20525] to CDA ab222515 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 16 kDa in Wild-type A549 cell lysates with no signal observed at this size in CDA knockout A549 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-CDA antibody [EPR20525] (<a href='/products/primary-antibodies/cda-antibody-epr20525-ab222515'>ab222515</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

CDA knockout A549 at 20 µg

Lane 2:

Western blot - Human CDA knockout A549 cell line (ab300883) at 20 µg

Lane 3:

Wild-type HCT at 20 µg

Lane 4:

CDA knockout HCT at 20 µg

Secondary

All lanes:

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

Predicted band size: 16 kDa

Observed band size: 16 kDa

false

关键信息

细胞类型

A549

种属

Human

组织

Lung

形式

Liquid

form

敲除验证

Western blot

突变描述

knockout

疾病

Carcinoma

产品详情

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.

规格

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性能和储存信息

基因名称
CDA
基因编辑类型
Knockout
基因编辑方法
CRISPR technology
敲除验证
Western blot
运输条件
Dry Ice
推荐的短期储存条件
-196°C
推荐的长期储存条件
-196°C

处理步骤

初始处理指南

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.

传代培养指南
  • 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.
培养基

F-12K + 10% FBS

低温储藏试剂

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

补充信息

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

Cytidine deaminase (CDA) also known as CDD is a zinc-dependent enzyme responsible for deaminating cytidine. The full form of CDA has a molecular mass of approximately 41 kDa. CDA is widely expressed in various tissues including liver kidney and leukocytes. It plays an important role in nucleotide metabolism by converting cytidine to uridine and 2'-deoxycytidine to 2'-deoxyuridine which impacts nucleic acid stability and synthesis. Researchers frequently use techniques such as ELISA and Western blotting to study CDA expression and activity often referencing it in contexts like "CDA Western" or "57 seconds CDA."
Biological function summary

CDA enzyme influences the pyrimidine salvage pathway by facilitating nucleotide degradation and recycling. It operates independently rather than as part of a protein complex. The activity of this enzyme ensures a balanced nucleotide pool essential for DNA replication and repair processes. This balance affects cellular proliferation and survival highlighting the importance of studying CDA protein in normal physiology and various pathological conditions.

Pathways

CDA contributes significantly to pyrimidine metabolism and its recycling pathway. It maintains a connection with ribonucleotide reductase which plays a role in the regulation of deoxyribonucleotide pools during DNA synthesis. This interplay highlights the enzyme’s involvement in DNA repair pathways ensuring proper cell division and genomic integrity. Within these pathways enzymes like cytidine triphosphate synthetase also relate to CDA by participating in nucleotide homeostasis.

Improper CDA function is linked to specific cancers including acute myeloid leukemia due to altered nucleotide metabolism leading to genomic instability. The enzyme's activity also affects the pharmacokinetics of cytidine analogues like gemcitabine used in cancer treatment. CDA overexpression might result in reduced drug efficacy thereby hindering therapy in affected patients. Connections exist between CDA and proteins involved in chemoresistance such as ribonucleotide reductase showing its significance in treatment outcomes.

细胞培养

生物安全等级

EU: 1 US: 1

贴壁/悬浮

Adherent

性别

Male

产品实验方案

Abcam Product Promise

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