CFH KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 3 and 1 bp deletion in exon 3 and 2 bp deletion in exon 3.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 3 and 1 bp deletion in exon 3 and 2 bp deletion in exon 3
Alternative names
AHUS 1, AMBP 1, ARMD 4, ARMS 1, CFAH_HUMAN, CFH, CFHL 3, Complement factor H, FH, Factor H, H factor 1, H factor 1 (complement), H factor 2 (complement), HF, HF 1, HF 2, HUS, MGC88246, adrenomedullin binding protein, age related maculopathy susceptibility 1, beta 1 H globulin, beta 1H, complement factor H, isoform b, factor H like 1
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CFH KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 3 and 1 bp deletion in exon 3 and 2 bp deletion in exon 3.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 10 bp deletion in exon 3 and 1 bp deletion in exon 3 and 2 bp deletion in exon 3
- Antibiotic resistance
- Puromycin 1µg/mL
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- CFH
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Male
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Factor H also known as complement factor H is an important regulatory protein in the complement system. It has a molecular mass of approximately 155 kDa. This protein is mainly expressed in the liver but it can also be found in low levels in other tissues. Factor H serves as a control element for complement activation particularly affecting the alternative pathway. It binds to C3b a central component of the complement system and accelerates the decay of C3 convertase as well as promotes the proteolytic inactivation of C3b by factor I.
- Biological function summary
Factor H limits the activity of the complement system to prevent damage to host tissues. The protein exists in the plasma in a soluble form. It functions by recognizing host cell surfaces via specific markers avoiding inappropriate activation. Factor H belongs to a group of proteins which include other regulators of complement activation. These proteins maintain the balance between effective immune defense and protection of host tissue from excessive immune responses.
- Pathways
Factor H is a part of the alternative complement pathway. This pathway is important for innate immune response involving proteins like factor P (properdin) which stabilizes C3 convertase. Factor H modulates these interactions to prevent unwarranted complement activity on host cells. Another related pathway is the classic complement pathway although factor H's involvement here is less direct since it primarily regulates the alternative pathway.
- Associated diseases and disorders
Factor H associations include atypical hemolytic uremic syndrome and age-related macular degeneration. Factor H deficiency or dysfunction can lead to uncontrolled complement activation resulting in kidney damage in atypical hemolytic uremic syndrome where it is also related to factor I. Additionally in age-related macular degeneration variants in the factor H gene are linked to increased susceptibility further highlighting the protein's importance in regulating immune responses.
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5 product images
Western blot - Human CFH (Factor H) knockout A549 cell line (ab267031)
Lanes 1-3: Merged signal (red and green). Green - Anti-Factor H antibody [EPR6225] ab133536 observed at 180 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-Factor H antibody [EPR6225] ab133536 Anti-Factor H antibody [EPR6225] was shown to specifically react with Factor H in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267031 (knockout cell lysate Human CFH (Factor H) knockout A549 cell lysate ab257150) was used. Wild-type and Factor H knockout samples were subjected to SDS-PAGE. Anti-Factor H antibody [EPR6225] ab133536 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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-Factor H antibody [EPR6225] (Anti-Factor H antibody [EPR6225] ab133536) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: CFH knockout A549 cell lysate at 20 µg
Lane 2: Western blot - Human CFH (Factor H) knockout A549 cell line (ab267031)
Lane 3: HaCaT cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 139 kDa
Observed band size: 180 kDa
Cell Culture - Human CFH (Factor H) knockout A549 cell line (ab267031)
Representative images of CFH knockout A549 cells, low and high confluency examples (top left and right respectively) and wild-type A549 cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
Sanger Sequencing - Human CFH (Factor H) knockout A549 cell line (ab267031)
Allele-2: 2 bp deletion in exon 3.
Sanger Sequencing - Human CFH (Factor H) knockout A549 cell line (ab267031)
Allele-1: 10 bp deletion in exon3
Sanger Sequencing - Human CFH (Factor H) knockout A549 cell line (ab267031)
Allele-3: 1 bp deletion in exon 3.
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