Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line
- Advanced Validation
- What is this?
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(1 Publication)
- WB
Lab
Western blot - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line (AB269503)
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 (<a href='/en-us/products/cell-lysates/human-hmox1-heme-oxygenase-1-knockout-a549-cell-lysate-ab269665'>ab269665</a>) 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
false
- WB
Lab
Western blot - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line (AB269503)
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 ° 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 (ab269503)
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
false
- NGS
Supplier Data
Next Generation Sequencing - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line (AB269503)
4 bp deletion after Val41 of the WT protein
- NGS
Supplier Data
Next Generation Sequencing - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell line (AB269503)
Knockout achieved by CRISPR/Cas9; X = 4 bp deletion; Frameshift : 100%
Complementary Products
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)](https://content.abcam.com/products/images/heme-oxygenase-1-antibody-ep1391y-ab52947--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img40284.jpg)
- 5
- 5
![Western blot - Anti-Nrf2 antibody [EP1808Y] (AB62352)](https://content.abcam.com/products/images/nrf2-antibody-ep1808y-ab62352--western-blot-img417623.jpg)
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
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.
Subculture guidelines
- 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.
Culture medium
F-12K + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
Experimental and therapeutic medicine 24:636 PubMed36160906
2022
Applications
Unspecified application
Species
Unspecified reactive species
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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