Human JAK2 knockout A549 cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human JAK2 knockout A549 cell line (AB267113)
Lanes 1- 4 : Merged signal (red and green). Green - ab108596 observed at 131 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab108596 was shown to react with JAK2 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267113 (knockout cell lysate ab256963) was used. Wild-type A549 and JAK2 knockout A549 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. ab108596 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-JAK2 antibody [EPR108(2)] (<a href='/en-us/products/primary-antibodies/jak2-antibody-epr1082-ab108596'>ab108596</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
JAK2 knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human JAK2 knockout A549 cell line (ab267113)
Lane 3:
K562 cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Predicted band size: 131 kDa
Observed band size: 131 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human JAK2 knockout A549 cell line (AB267113)
Allele-1 : 5 bp deletion in exon3
- Sanger seq
Unknown
Sanger Sequencing - Human JAK2 knockout A549 cell line (AB267113)
Allele-2 : 1 bp insertion in exon 3.
Complementary Products
Primary Antibodies
AB32101
Anti-JAK2 (phospho Y1007 + Y1008) + JAK1 (phospho Y1034 + Y1035) antibody [E132]
20ul selling size|RabMAb|Recombinant
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK2 (phospho Y1007 + Y1008) + JAK1 (phospho Y1034 + Y1035) antibody [E132] (AB32101)](https://content.abcam.com/products/images/jak2-phospho-y1007-y1008-jak1-phospho-y1034-y1035-antibody-e132-ab32101--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img179905.jpg)
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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
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
JAK2 is important for transmitting signals that dictate cell growth survival and differentiation within the hematopoietic system. It operates bodily functions by forming complexes with specific phosphorylation sites on its associated receptors. Through this formation JAK2 influences the signaling cascade particularly by interacting with other signal transducers and activators where it phosphorylates and becomes activated. This action affects gene transcription directly correlated with cellular proliferation and differentiation processes.
Pathways
The function of JAK2 integrates into the JAK-STAT signaling pathway which is an important pathway in the regulation of immune function growth and development. It works in conjunction with proteins such as STAT3 and STAT5 to transmit signals from cytokine receptors to the nucleus. This pathway critically impacts responses like inflammation and hematopoiesis aligning with its role in precursor proliferation within the bone marrow and various immune cells' function.
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
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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