Human CFH (Factor H) knockout A549 cell line
- Advanced Validation
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CFH KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. 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. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View 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
- WB
Lab
Western blot - Human CFH (Factor H) knockout A549 cell line (AB267031)
Lanes 1-3 : Merged signal (red and green). Green - ab133536 observed at 180 kDa. Red - loading control ab8245 observed at 36 kDa.
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 ab257150) was used. Wild-type and Factor H knockout samples were subjected to SDS-PAGE. ab133536 and 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 (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-Factor H antibody [EPR6225] (<a href='/en-us/products/primary-antibodies/factor-h-antibody-epr6225-ab133536'>ab133536</a>) 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
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: 139 kDa
Observed band size: 180 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CFH (Factor H) knockout A549 cell line (AB267031)
Allele-2 : 2 bp deletion in exon 3.
- Sanger seq
Unknown
Sanger Sequencing - Human CFH (Factor H) knockout A549 cell line (AB267031)
Allele-3 : 1 bp deletion in exon 3.
- Cell Culture
Unknown
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 seq
Unknown
Sanger Sequencing - Human CFH (Factor H) knockout A549 cell line (AB267031)
Allele-1 : 10 bp deletion in exon3
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
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.
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
Complementary Products
Primary Antibodies
AB305137
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Primary Antibodies
AB259716
Anti-Alpha 2 Antiplasmin antibody [EPR22550-263] - BSA and Azide free (Detector)
RabMAb|Recombinant
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Product promise
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