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AB269665

Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate

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HMOX1 KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift: 100%.
3 Images
Western blot - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate (AB269665)
  • WB

Lab

Western blot - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate (AB269665)

Lane 1 : Human wild-type A549 cell lysate (20 ug)
Lane 2 : Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysatecell lysate (20 ug)
Lane 3 : Human spleen tissue lysate (20 ug)
Lane 4 : HL-60 cell lysate (20 ug)
Lane 5 : MCF7 cell lysate (20 ug)
Lane 6 : HeLa cell lysate (20 ug)
Lane 7 : A549 cell lysate (20 ug)

Lanes 1 - 7 : Merged signal (red and green). Green - ab52947 observed at 33 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab52947 was shown to react with Heme Oxygenase 1 in wild-type A549 cells in Western blot with loss of signal observed in HMOX1 knockout cell line ab269503 (knockout cell lysate ab269665). Wild-type A549 and HMOX1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab52947 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Heme Oxygenase 1 antibody [EP1391Y] (<a href='/en-us/products/primary-antibodies/heme-oxygenase-1-antibody-ep1391y-ab52947'>ab52947</a>) at 1/2000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

HMOX1 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-hmox1-heme-oxygenase-1-knockout-a549-cell-line-ab269503'>ab269503</a>)

Lane 3:

Human Spleen tissue lysate at 20 µg

Lane 4:

HL-60 cell lysate at 20 µg

Lane 5:

MCF7 cell lysate at 20 µg

Lane 6:

HeLa cell lysate at 20 µg

Lane 7:

A549 cell lysate at 20 µg

Predicted band size: 33 kDa

Observed band size: 33 kDa

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Next Generation Sequencing - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate (AB269665)
  • NGS

Supplier Data

Next Generation Sequencing - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate (AB269665)

Knockout achieved by CRISPR/Cas9; X = 4 bp deletion; Frameshift : 100%

Western blot - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate (AB269665)
  • WB

Lab

Western blot - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate (AB269665)

Lanes 1 - 7 : Merged signal (red and green). Green - ab68477 observed at 33 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab68477 was shown to react with Heme Oxygenase 1 in wild-type A549 cells in Western blot with loss of signal observed in HMOX1 knockout cell line ab269503 (knockout cell lysate ab269665). Wild-type A549 and HMOX1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab68477 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Heme Oxygenase 1 antibody [EPR1390Y] (<a href='/en-us/products/primary-antibodies/heme-oxygenase-1-antibody-epr1390y-ab68477'>ab68477</a>) at 1/10000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Western blot - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate (ab269665) at 20 µg

Lane 3:

Human Spleen tissue lysate at 20 µg

Lane 4:

HL-60 cell lysate at 20 µg

Lane 5:

MCF7 cell lysate at 20 µg

Lane 6:

HeLa cell lysate at 20 µg

Lane 7:

A549 cell lysate at 20 µg

Predicted band size: 33 kDa

Observed band size: 33 kDa

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Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 4 bp deletion Frameshift: 100%

Disease

Carcinoma

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Complementary Products

Cell Lines & Lysates

AB269503

Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line

Advanced Validation

Western blot - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line (AB269503)

  • 0
  • 0
Primary Antibodies

AB52947

Anti-Heme Oxygenase 1 antibody [EP1391Y]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Heme Oxygenase 1 antibody [EP1391Y] (AB52947)

  • 5
  • 5
Primary Antibodies

AB128936

Anti-Syndecan-1 antibody [EPR6454]

20ul selling size|RabMAb|Recombinant

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syndecan-1 antibody [EPR6454] (AB128936)

  • 5
  • 7
Secondary Antibodies

AB150113

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

Immunocytochemistry/ Immunofluorescence - Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (AB150113)

  • 5
  • 15
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Reactivity data

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

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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab269665-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human HMOX1 knockout A549 cell lysate", "number":"AB269665-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type A549 cell lysate", "number":"AB269665-CMP01", "productcode":"" } ] } } }
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Properties and storage information

Gene name
HMOX1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation 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.

Heme Oxygenase 1 also known as HO-1 or HMOX1 is an enzyme that plays an important mechanistic role in heme catabolism. It catalyzes the degradation of heme into biliverdin carbon monoxide and free iron. This process involves the cleavage of the heme ring. HO-1 has a molecular weight of approximately 32 kDa. It is widely expressed in numerous tissues but is especially abundant in the liver and spleen. Its expression is induced by heme and other stress stimuli such as heavy metals cytokines and reactive oxygen species.
Biological function summary

Heme Oxygenase 1 serves important protective functions in the body. It is not part of a larger complex but its products such as carbon monoxide and biliverdin have their own biological activities. Carbon monoxide produced by HO-1 has antiflammatory properties and can modulate apoptotic pathways. Biliverdin is reduced to bilirubin which acts as an antioxidant. The enzyme therefore directly influences cellular stress responses and maintains cellular homeostasis through these processes.

Pathways

Heme Oxygenase 1 is integrally involved in oxidative stress response and heme metabolism. It participates in the cellular response to oxidative damage by reducing oxidative stress and promoting cytoprotection. Through its heme degradation activity it is connected with the synthesis of biologically active molecules like bilirubin and carbon monoxide. Heme Oxygenase 1 activity is related to other proteins in oxidative stress pathways such as Nuclear Factor Erythroid 2-Related Factor 2 (Nrf2) which regulates its expression and globins which are sources of heme for HO-1 activity.

Heme Oxygenase 1 has been linked to conditions like cardiovascular diseases and neurodegenerative disorders. Its expression can attenuate the severity of atherosclerosis where oxidative stress is an important factor. In neurodegenerative diseases HO-1’s antioxidant properties may provide neuroprotection by mitigating oxidative damage. The protein's interactions with inflammatory cytokines such as Interleukin-6 and tumor necrosis factor-alpha influence its activity in these disease contexts.
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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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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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