Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate (ab269665)
Overview
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Product name
Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate
See all Heme Oxygenase 1 kits -
Product overview
Knockout cell lysate achieved by CRISPR/Cas9.
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Parental Cell Line
A549 -
Organism
Human -
Mutation description
Knockout achieved by CRISPR/Cas9; X = 4 bp deletion; Frameshift: 100% -
Passage number
<20 -
Knockout validation
Next Generation Sequencing (NGS), Western Blot (WB) -
Reconstitution notes
To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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Notes
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 - found here. 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.
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.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.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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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -80°C. Please refer to protocols. -
Components 1 kit ab280540 - Human HMOX1 knockout A549 cell lysate 1 x 100µg ab259782 - Human wild-type A549 cell lysate 1 x 100µg -
Research areas
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Cell type
epithelial -
Disease
Carcinoma -
Gender
Male
Target
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Function
Heme oxygenase cleaves the heme ring at the alpha methene bridge to form biliverdin. Biliverdin is subsequently converted to bilirubin by biliverdin reductase. Under physiological conditions, the activity of heme oxygenase is highest in the spleen, where senescent erythrocytes are sequestrated and destroyed. -
Sequence similarities
Belongs to the heme oxygenase family. -
Cellular localization
Microsome. Endoplasmic reticulum. - Information by UniProt
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Alternative names
- 32 kD
- bK286B10
- D8Wsu38e
see all
Associated products
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab269665 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration. Predicted molecular weight: 33 kDa.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. Predicted molecular weight: 33 kDa. |
Images
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Next Generation Sequencing - Human HMOX1 (Heme Oxygenase 1) knockout A549 cell lysate (ab269665)Knockout achieved by CRISPR/Cas9; X = 4 bp deletion; Frameshift: 100%
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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 - 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.
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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.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab269665 has not yet been referenced specifically in any publications.