Human SYK knockout THP-1 cell line
- Advanced Validation
- What is this?
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(0 Publication)
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human SYK knockout THP-1 cell line (AB288700)
ab244968 was shown to react with SYK in wild-type THP-1 cells in immunocytochemistry with loss of signal observed in a SYK knockout cell line. ab244968 at 1/1000 dilution.
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
- WB
Lab
Western blot - Human SYK knockout THP-1 cell line (AB288700)
Western blot : Anti-SYK antibody [EP573Y] (ab40781) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab40781 was shown to bind specifically to SYK. A band was observed at 72 kDa in wild-type THP-1 cell lysates with no signal observed at this size in SYK knockout cell line ab288700 (knockout cell lysate ab289593). To generate this image, wild-type and SYK knockout THP-1 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.
All lanes:
Western blot - Anti-Syk antibody [EP573Y] (<a href='/en-us/products/primary-antibodies/syk-antibody-ep573y-ab40781'>ab40781</a>) at 1/1000 dilution
Lanes 1 - 4:
Western blot at 20 µg
Lane 2:
Western blot - Human SYK knockout THP-1 cell line (ab288700)
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Predicted band size: 72 kDa
Observed band size: 72 kDa
false
- WB
Lab
Western blot - Human SYK knockout THP-1 cell line (AB288700)
ab244968 was shown to react with SYK in wild-type THP-1 cells in Western blot with loss of signal observed in SYK knockout cell line ab288700. Wild-type THP-1 and SYK knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab244968 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.
These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes:
Western blot - Anti-Syk antibody [EPR573-69] - BSA and Azide free (Detector) (<a href='/en-us/products/primary-antibodies/syk-antibody-epr573-69-bsa-and-azide-free-detector-ab244968'>ab244968</a>) at 1/1000 dilution
Lane 1:
Wild-type THP-1 lysate at 30 µg
Lane 2:
SYK knock-out THP-1 lysate at 30 µg
false
- Sanger seq
Supplier Data
Sanger Sequencing - Human SYK knockout THP-1 cell line (AB288700)
127 bp deletion in exon 2.
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human SYK knockout THP-1 cell line (AB288700)
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human SYK knockout THP-1 cell line (AB288700)
- ICC/IF
Lab
Immunocytochemistry/ Immunofluorescence - Human SYK knockout THP-1 cell line (AB288700)
- WB
Lab
Western blot - Human SYK knockout THP-1 cell line (AB288700)
All lanes:
Western blot - Anti-Syk antibody [EP573Y] (<a href='/en-us/products/primary-antibodies/syk-antibody-ep573y-ab40781'>ab40781</a>) at 1/1000 dilution
Lane 1:
Wild-type THP-1 lysate at 20 µg
Lane 2:
Western blot - Human SYK knockout THP-1 cell line (ab288700) at 20 µg
Observed band size: 72 kDa
false
- WB
Lab
Western blot - Human SYK knockout THP-1 cell line (AB288700)
All lanes:
Western blot - Anti-Syk antibody [EPR19414-176] - BSA and Azide free (Capture) (<a href='/en-us/products/primary-antibodies/syk-antibody-epr19414-176-bsa-and-azide-free-capture-ab244701'>ab244701</a>) at 1/1000 dilution
Lane 1:
Wild-type THP-1 lysate at 20 µg
Lane 2:
Western blot - Human SYK knockout THP-1 cell line (ab288700) at 20 µg
Observed band size: 72 kDa
false
- WB
Lab
Western blot - Human SYK knockout THP-1 cell line (AB288700)
All lanes:
Western blot - Anti-Syk antibody [SYK-01] (<a href='/en-us/products/primary-antibodies/syk-antibody-syk-01-ab3993'>ab3993</a>) at 1/500 dilution
Lane 1:
Wild-type THP-1 lysate at 20 µg
Lane 2:
Western blot - Human SYK knockout THP-1 cell line (ab288700) at 20 µg
Observed band size: 72 kDa
false
Complementary Products
Primary Antibodies
AB62350
Anti-Syk (phospho Y525) antibody [EP575(2)Y]
20ul selling size|RabMAb|Recombinant
![Western blot - Anti-Syk (phospho Y525) antibody [EP575(2)Y] (AB62350)](https://content.abcam.com/products/images/syk-phospho-y525-antibody-ep5752y-ab62350--western-blot-img438263.jpg)
- 0
- 0
![Western blot - Anti-Lyn antibody [Y497] (AB32398)](https://content.abcam.com/products/images/lyn-antibody-y497-ab32398--western-blot-img159428.jpg)
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
Zygosity
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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x105 cells/mL.Small amounts of fresh media can be added until cell number/viability improves.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
- It is not recommended to allow the cell density to exceed 1x106 cells/mL.
Culture medium
RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
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
Syk plays a role in transmitting signals from the cell surface to the interior of the cell facilitating various cellular activities. It is a part of signal transduction complexes and functions in the regulation of immune cell activation differentiation and survival. It affects processes like Fc receptor signaling in macrophages and neutrophils impacting immune responses and inflammation. The phospho-Syk form influences the strength and duration of signaling affecting various cellular functions.
Pathways
Syk is significant in the B-cell receptor (BCR) signaling pathway and Fc receptor signaling pathway. Within these pathways Syk interacts with proteins like Lyn and Fyn which are other kinases involved in BCR signal transduction. Syk phosphorylation noted as p-Syk plays a part in activating downstream molecules such as phospholipase C gamma (PLCγ) to mediate cellular responses to external stimuli. The critical role of Syk in these pathways influences both adaptive and innate immune responses.
Quality control
STR analysis
VWA, TPOX, CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
Gender
Male
Target data
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