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AB257143

Human ASS1 knockout HeLa cell lysate

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ASS1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and Insertion of the selection cassette in exon 3.
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Western blot - Human ASS1 knockout HeLa cell lysate (AB257143)
  • WB

Lab

Western blot - Human ASS1 knockout HeLa cell lysate (AB257143)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : ASS1 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab170900 observed at 47 kDa. Red - loading control ab8245 observed at 37 kDa.

ab170900 Anti-ASS1 antibody [EPR12399(B)] - C-terminal was shown to specifically react with ASS1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264989 (knockout cell lysate ab257143) was used. Wild-type and ASS1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab170900 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 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ASS1 antibody [EPR12399(B)] - C-terminal (<a href='/en-us/products/primary-antibodies/ass1-antibody-epr12399b-c-terminal-ab170900'>ab170900</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ASS1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ASS1 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ass1-knockout-hela-cell-line-ab264989'>ab264989</a>)

Predicted band size: 47 kDa

Observed band size: 47 kDa

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Western blot - Human ASS1 knockout HeLa cell lysate (AB257143)
  • WB

Lab

Western blot - Human ASS1 knockout HeLa cell lysate (AB257143)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : ASS1 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab170952 observed at 47 kDa. Red - loading control ab8245 observed at 37 kDa.

ab170952 Anti-ASS1 antibody [EPR12398] was shown to specifically react with ASS1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264989 (knockout cell lysate ab257143) was used. Wild-type and ASS1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab170952 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 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ASS1 antibody [EPR12398] (<a href='/en-us/products/primary-antibodies/ass1-antibody-epr12398-ab170952'>ab170952</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ASS1 knockout HeLa cell lysate at 20 µg

Predicted band size: 47 kDa

Observed band size: 47 kDa

false

Sanger Sequencing - Human ASS1 knockout HeLa cell lysate (AB257143)
  • Sanger seq

Unknown

Sanger Sequencing - Human ASS1 knockout HeLa cell lysate (AB257143)

Allele-2 : Insertion of the selection cassette in exon 3

Sanger Sequencing - Human ASS1 knockout HeLa cell lysate (AB257143)
  • Sanger seq

Unknown

Sanger Sequencing - Human ASS1 knockout HeLa cell lysate (AB257143)

Allele-1 : 1 bp insertion in exon 3

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 3 and Insertion of the selection cassette in exon 3.

Disease

Adenocarcinoma

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Complementary Products

Primary Antibodies

AB170952

Anti-ASS1 antibody [EPR12398]

RabMAb|Recombinant|KO Validated|20ul selling size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ASS1 antibody [EPR12398] (AB170952)

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  • 1
Proteins & Peptides

AB286071

Recombinant E. coli ASS1 protein (Active)

Functional Studies - Recombinant E. coli ASS1 protein (Active) (AB286071)

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  • 0
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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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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What's included?

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Properties and storage information

Gene name
ASS1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

ASS1 or argininosuccinate synthase 1 is an enzyme that plays an important role in the urea cycle. It catalyzes the conversion of citrulline and aspartate into argininosuccinate using ATP in the process. The ASS1 protein is commonly expressed in the liver where it actively participates in the detoxification of ammonia. It has a molecular mass of approximately 46 kDa. Some alternate names for this protein include ASS and ASS-1.
Biological function summary

Argininosuccinate synthase 1 contributes significantly to the synthesis of arginine an essential amino acid through its conversion activities. The enzyme operates as a homotrimer a complex consisting of three identical subunits. This enzymatic activity is indispensable in maintaining the balance of nitrogen and ammonia within the body particularly important for liver and kidney functions.

Pathways

ASS1 is a critical component of both the urea cycle and the nitric oxide metabolism pathway. The urea cycle helps in detoxifying ammonia by converting it into urea which is excreted in urine. In nitric oxide metabolism ASS1 links with nitric oxide synthase to facilitate the use of arginine in producing nitric oxide a vital signaling molecule. The pathway engagement partners include proteins like citrin which supplies aspartate for the reaction.

ASS1 has implications in citrullinemia and hepatocellular carcinoma. Citrullinemia results from mutations in the ASS1 gene leading to defective urea cycle function and ammonia accumulation. In cancer particularly hepatocellular carcinoma low expression of ASS1 correlates with poor prognosis as cancer cells often exhibit arginine auxotrophy. Connections through these conditions involve interactions with enzymes such as arginase which also influences arginine availability.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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