Human AIFM1 (AIF) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(1 Publication)
AIFM1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon 2 and 23 bp deletion in exon 2 and 2 bp insertion in exon 2. 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
PDCD8, Programmed cell death protein 8, AIF, AIFM1
- WB
Lab
Western blot - Human AIFM1 (AIF) knockout HEK-293T cell line (AB266347)
Lanes 1- 2 : Merged signal (red and green). Green - ab32516 observed at 67 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32516 was shown to react with AIF in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266347 (knockout cell lysate ab256834) was used. Wild-type HEK-293T and AIFM1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32516 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4° at a 1 in 1000 dilution and a 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-AIF antibody [E20] - Mitochondrial Marker (<a href='/en-us/products/primary-antibodies/aif-antibody-e20-mitochondrial-marker-ab32516'>ab32516</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
AIFM1 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human AIFM1 (AIF) knockout HEK-293T cell line (ab266347)
Predicted band size: 67 kDa
Observed band size: 67 kDa
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- Cell Culture
Lab
Cell Culture - Human AIFM1 (AIF) knockout HEK-293T cell line (AB266347)
Representative images AIFM1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human AIFM1 (AIF) knockout HEK-293T cell line (AB266347)
Allele-2 : 1 bp insertion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human AIFM1 (AIF) knockout HEK-293T cell line (AB266347)
Allele-3 : 2 bp insertion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human AIFM1 (AIF) knockout HEK-293T cell line (AB266347)
Allele-1 : 23 bp deletion in exon 2
Complementary Products
Primary Antibodies
AB196847
Alexa Fluor® 647 Anti-AIF antibody [E20] - Mitochondrial Marker
RabMAb|Recombinant
![Flow Cytometry (Intracellular) - Alexa Fluor® 647 Anti-AIF antibody [E20] - Mitochondrial Marker (AB196847)](https://content.abcam.com/products/images/alexa-fluor-647-aif-antibody-e20-mitochondrial-marker-ab196847--flow-cytometry-intracellular-img461599.jpg)
- 0
- 0
Cell Lines & Lysates
AB266272
Human JUP (gamma Catenin) knockout HEK-293T cell line
Advanced Validation
- 0
- 0
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
AIF functions as an important element in mitochondrial-mediated apoptosis. Once released from mitochondria AIF triggers a caspase-independent pathway of apoptosis making it distinct from other pathways where caspases are central. While not part of a larger protein complex AIF interacts closely with several mitochondrial and nuclear elements to execute its functions. Researchers often monitor AIF activity using apoptosis ELISA kits which help in precise detection and quantification of this protein within cellular systems.
Pathways
The presence and activity of AIF align with the broader apoptotic pathway and mitochondrial respiration. AIF links significantly with other molecules like cytochrome c released during apoptosis although cytochrome c follows a caspase-dependent pathway. It also shows involvement in various cellular injury and stress response pathways reinforcing its role as a major player in cellular fate decisions and intrinsic apoptotic mechanisms.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
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.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Target data
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
Nature communications 16:10817 PubMed41326385
2025
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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