JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB257348

Human ALAS1 knockout HEK-293T cell lysate

Be the first to review this product! Submit a review

|

(0 Publication)

ALAS1 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.

View Alternative Names

5 aminolevulinate synthase nonspecific mitochondrial, 5 aminolevulinic acid synthase, 5-aminolevulinate synthase, 5-aminolevulinic acid synthase 1, ALAS, ALAS 3, ALAS HOUSEKEEPING TYPE, ALAS N, ALAS-H, Alas 1, Aminolevulinate delta synthase 1, Aminolevulinic acid synthase 1, Delta ALA synthetase, Delta aminolevulinate synthase, Delta-ALA synthase 1, Delta-aminolevulinate synthase 1, HEM1_HUMAN, MIG 4, Migration inducing protein 4, mitochondrial, nonspecific

3 Images
Western blot - Human ALAS1 knockout HEK-293T cell lysate (AB257348)
  • WB

Unknown

Western blot - Human ALAS1 knockout HEK-293T cell lysate (AB257348)

Lane 1 : Wild-type HeLa cell lysate (20 ug)
Lane 2 : ALAS1 knockout HeLa cell lysate (20 ug)
Lane 3 : HepG2 cell lysate (20 ug)
Lane 4 : HEK-293 cell lysate (20 ug)

ab154860 was shown to specifically react with Alas1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab266473 (knockout cell lysate ab257348) was used. Wild-type and Alas1 knockout samples were subjected to SDS-PAGE. ab154860 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4oC 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-Alas1 antibody [EPR10247] - Mitochondrial Marker (<a href='/en-us/products/primary-antibodies/alas1-antibody-epr10247-mitochondrial-marker-ab154860'>ab154860</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ALAS1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ALAS1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-alas1-knockout-hek-293t-cell-line-ab266473'>ab266473</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

HEK-293 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 71 kDa

Observed band size: 71 kDa

false

Sanger Sequencing - Human ALAS1 knockout HEK-293T cell lysate (AB257348)
  • Sanger seq

Unknown

Sanger Sequencing - Human ALAS1 knockout HEK-293T cell lysate (AB257348)

Allele-1 : 1 bp insertion in exon 3

Sanger Sequencing - Human ALAS1 knockout HEK-293T cell lysate (AB257348)
  • Sanger seq

Unknown

Sanger Sequencing - Human ALAS1 knockout HEK-293T cell lysate (AB257348)

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

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

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.

style
left-column

Reactivity data

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

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.

style
left-column

What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257348-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human ALAS1 knockout HEK293T cell lysate", "number":"AB257348-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB257348-CMP02", "productcode":"" } ] } } }
style
left-column

Properties and storage information

Gene name
ALAS1
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
style
left-column

Supplementary information

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

'Alas1' also known as 5-Aminolevulinic acid synthase 1 is a mitochondrial enzyme with a mass of approximately 64 kDa. This enzyme plays an important role in heme biosynthesis catalyzing the first step in the pathway by converting glycine and succinyl-CoA to 5-aminolevulinic acid. Unfortunately it does not function alone and often requires the presence of pyridoxal phosphate as a cofactor. 'Alas1' is ubiquitously expressed in a variety of tissues including liver and bone marrow where heme production is critical.
Biological function summary

'Alas1' serves as a rate-limiting enzyme in the heme synthesis pathway. Being the first step it sets the pace for the entire process. It does not form part of any larger protein complex and operates with its required cofactors. Beyond metabolic roles its regulation influences several cellular processes ensuring heme availability precisely matches cellular demand.

Pathways

'Alas1' performs a fundamental role in the mitochondrial heme biosynthesis pathway. This pathway is essential for synthesizing heme a component critical to various cellular functions such as oxygen transport and electron transfer. 'Alas1' interacts with proteins like ferrochelatase the enzyme completing the heme biosynthesis pathway. Coordination between 'Alas1' and other proteins ensures efficient production of heme enabling proper cellular function and adaptation to changes in cellular and systemic conditions.

'Alas1' is involved in conditions such as acute intermittent porphyria and certain anemias. Its dysregulation can lead to impaired heme production with downstream effects on cell metabolism and organ function. 'Alas1' has been shown to interact with HMBS (hydroxymethylbilane synthase) a protein linked to different types of porphyria. Understanding its role in these conditions could lead to targeted therapies that modulate its activity to alleviate symptoms or correct underlying biochemical dysfunctions.
style
left-column

Quality control

STR analysis

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

style
left-column

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

style
left-column

Product protocols

style
left-column
style
left-column
style
right-column

Complementary Products

Proteins & Peptides

AB259412

Recombinant human VEGF 165A protein (Active)

Premium Bioactive

Mass Spectrometry - Recombinant human VEGF 165A protein (Active) (AB259412)

  • 0
  • 0
Proteins & Peptides

AB180317

Recombinant Human POLR1C protein

SDS-PAGE - Recombinant Human POLR1C protein (AB180317)

  • 0
  • 0
Primary Antibodies

AB154860

Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker

RabMAb|Recombinant|Lab Essentials|KO Validated

Western blot - Anti-Alas1 antibody [EPR10247] - Mitochondrial Marker (AB154860)

  • 5
  • 1
Primary Antibodies

AB186130

Anti-SHMT1 antibody

Western blot - Anti-SHMT1 antibody (AB186130)

  • 0
  • 0

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'.

For licensing inquiries, please contact partnerships@abcam.com