Human TAB2 knockout A549 cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human TAB2 knockout A549 cell line (AB301263)
Western blot : Anti-TAB2 antibody staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, the antibody was shown to bind specifically to TAB2. A band was observed at 80 kDa in wild-type A549 cell lysates with no signal observed at this size in TAB2 knockout cell line ab301263. To generate this image, wild-type and TAB2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
A549 cell lysate at 20 µg
Lane 3:
Raji cell lysate at 20 µg
Lane 4:
HEK-293 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
false
- NGS
Lab
Next Generation Sequencing - Human TAB2 knockout A549 cell line (AB301263)
64 bp deletion after Thr 179 (allele 1); 55 bp deletion after Ala 181 (allele 2) of WT protein
- WB
Lab
Western blot - Human TAB2 knockout A549 cell line (AB301263)
Western blot : Anti-TAB2 antibody [EPR19670-161] ab317602 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 80 kDa in Wild-type A549 cell lysates with no signal observed at this size in TAB2 knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween™ 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-TAB2 antibody [EPR19670-161] (<a href='/en-us/products/primary-antibodies/tab2-antibody-epr19670-161-ab317602'>ab317602</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 whole cell lysate at 20 µg
Lane 2:
Western blot - Human TAB2 knockout A549 cell line (ab301263) at 20 µg
Lane 3:
HEK-293 whole cell lysate at 20 µg
Lane 4:
Raji whole cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 76 kDa
Observed band size: 80 kDa
false
Complementary Products
Primary Antibodies
AB317603
Anti-TAB2 antibody [EPR19670-161] - BSA and Azide free
Recombinant|RabMAb
![Immunoprecipitation - Anti-TAB2 antibody [EPR19670-161] - BSA and Azide free (AB317603)](https://content.abcam.com/products/images/tab2-antibody-epr19670-161-bsa-and-azide-free-ab317603--immunoprecipitation-img431955.jpg)
- 0
- 0
Reactivity data
Product details
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
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
TAB2 maintains a significant position in cellular signaling as part of larger protein complexes. It associates with TAK1 (transforming growth factor-beta-activated kinase 1) to regulate NF-kappaB and MAPK pathways. Through these interactions TAB2 helps cells respond to environmental signals mediating transcriptional responses necessary for adaptation and defense mechanisms. In particular TAB2's role in these complexes underlines its importance in immune response modulation and cellular homeostasis.
Pathways
TAB2 is deeply involved in both the NF-kappaB and MAPK signaling pathways where it cooperates intimately with proteins such as TAK1 and TAB1. These pathways are essential for gene expression regulation in response to pro-inflammatory signals. TAB2 facilitates the activation of IKK complex within the NF-kappaB pathway and JNK in the MAPK pathway. By influencing these pathways TAB2 governs important cellular processes like apoptosis proliferation and differentiation.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
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