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AB261809

人CPS1 knockout HeLa cell line

Human CPS1 knockout HeLa cell line

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CPS1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 15 and Insertion of the selection cassette in exon 15. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.

查看别名

CPSM_HUMAN, CPSase 1, CPSase I, Carbamoyl phosphate synthase, Carbamoyl phosphate synthase [ammonia] mitochondrial, Carbamoyl phosphate synthetase 1, Carbamoyl phosphate synthetase 1 mitochondrial, Carbamoyl-phosphate synthase [ammonia], Carbamoyl-phosphate synthetase I, MS738, mitochondrial

4 Images
Western blot - Human CPS1 knockout HeLa cell line (AB261809)
  • WB

Lab

Western blot - Human CPS1 knockout HeLa cell line (AB261809)

Lanes 1-3 : Merged signal (red and green). Green - ab155083 observed at 165 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab155083 Anti-CPS1 antibody [EPR7493-29] was shown to specifically react with CPS1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261809 (knockout cell lysate ab257121) was used. Wild-type and CPS1 knockout samples were subjected to SDS-PAGE. ab155083 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CPS1 antibody [EPR7493-29] (<a href='/products/primary-antibodies/cps1-antibody-epr7493-29-ab155083'>ab155083</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CPS1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CPS1 knockout HeLa cell line (ab261809)

Lane 3:

Human liver tissue lysate at 20 µg

Predicted band size: 165 kDa

Observed band size: 165 kDa

false

Western blot - Human CPS1 knockout HeLa cell line (AB261809)
  • WB

Lab

Western blot - Human CPS1 knockout HeLa cell line (AB261809)

Lanes 1-3 : Merged signal (red and green). Green - ab129076 observed at 165 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab129076 Anti-CPS1 antibody [EPR7493-3] was shown to specifically react with CPS1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261809 (knockout cell lysate ab257121) was used. Wild-type and CPS1 knockout samples were subjected to SDS-PAGE. ab129076 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CPS1 antibody [EPR7493-3] (<a href='/products/primary-antibodies/cps1-antibody-epr7493-3-ab129076'>ab129076</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CPS1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CPS1 knockout HeLa cell line (ab261809)

Lane 3:

Human liver tissue lysate at 20 µg

Predicted band size: 165 kDa

Observed band size: 165 kDa

false

Sanger Sequencing - Human CPS1 knockout HeLa cell line (AB261809)
  • Sanger seq

Unknown

Sanger Sequencing - Human CPS1 knockout HeLa cell line (AB261809)

Allele-2 : Insertion of the selection cassette in exon 15.

Sanger Sequencing - Human CPS1 knockout HeLa cell line (AB261809)
  • Sanger seq

Unknown

Sanger Sequencing - Human CPS1 knockout HeLa cell line (AB261809)

Allele-1 : 1 bp insertion in exon 15.

关键信息

细胞类型

HeLa

种属

Human

组织

Cervix

形式

Liquid

form

敲除验证

Sanger Sequencing,Western blot

突变描述

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 15 and Insertion of the selection cassette in exon 15

疾病

Adenocarcinoma

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反应性数据

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

We will provide viable cells that proliferate on revival.

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

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规格

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性能和储存信息

基因名称
CPS1
基因编辑类型
Knockout
基因编辑方法
CRISPR technology
敲除验证
Sanger Sequencing, 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
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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.
培养基

DMEM (High Glucose) + 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.

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补充信息

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

CPS1 also known as carbamoyl-phosphate synthetase 1 is an important mitochondrial enzyme with a mass of around 165 kDa. It acts in the urea cycle by converting ammonia and bicarbonate into carbamoyl phosphate. CPS1 expression occurs in the liver and intestinal enterocytes which situates it in tissues responsible for metabolism and detoxification processes. This enzyme plays a critical role in the removal of excess nitrogen from the body.
Biological function summary

Carbamoyl-phosphate synthetase 1 functions as an initial step enzyme in the urea cycle which is a metabolic pathway for nitrogen disposal. It does not function alone as it forms part of multi-enzyme complexes to enhance efficiency in the cycle. These complexes facilitate the process of converting toxic ammonia into urea which the body can safely excrete. The proper function of CPS1 ensures that ammonia levels remain non-toxic.

Pathways

The urea cycle remains the chief pathway involving CPS1 collaborating with enzymes like ornithine transcarbamylase and argininosuccinate synthetase. This cycle intersects with the mitochondrial respiratory chain showcasing a relationship between CPS1 and cellular energy balance. Besides the urea cycle the enzyme also links to other metabolic functions like gluconeogenesis given its prerequisite for maintaining a stable nitrogen homeostasis.

CPS1 deficiencies can lead to urea cycle disorders like hyperammonemia characterized by elevated ammonia levels in blood. This may result in neurological damage if untreated. Another condition related to CPS-1 is pulmonary arterial hypertension where studies suggest mishandling of ammonia may alter blood pressure regulation. Genetic mutations affecting the CPS1 protein often underlie these conditions highlighting the importance of genetic screening in affected individuals.
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质量控制

STR 分析

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

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细胞培养

生物安全等级

EU: 2 US: 2

贴壁/悬浮

Adherent

性别

Female

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产品实验方案

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靶点信息

See full target information CPS1
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搭配使用产品

Primary Antibodies

AB129076

Anti-CPS1 antibody [EPR7493-3]

RabMAb|Recombinant|KO Validated|20ul selling size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CPS1 antibody [EPR7493-3] (AB129076)

  • 0
  • 0
Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20
Multiplex Kits

AB197478

Non-Human Primate Cytokine Antibody Array (10 Targets) - Quantitative

Multiplex Protein Detection - Non-Human Primate Cytokine Antibody Array (10 Targets) - Quantitative (AB197478)

  • 0
  • 0
Primary Antibodies

AB15580

Anti-Ki67 antibody

BOND RX™ Validated|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody (AB15580)

  • 5
  • 230

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