Mouse ARC knockout Neuro-2a cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Mouse ARC knockout Neuro-2a cell line (AB280071)
False colour image of Western blot : Anti-Arc antibody [EPR18950] staining at 1/500 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot ab183183 was shown to bind specifically to Arc. A band was observed at 45 kDa in wild-type Neuro-2a cell lysates with no signal observed at this size in arc knockout cell line ab280071 (knockout cell lysate ab280130). To generate this image wild-type and arc knockout Neuro-2a cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4°. 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-Arc antibody [EPR18950] (<a href='/en-us/products/primary-antibodies/arc-antibody-epr18950-ab183183'>ab183183</a>) at 1/500 dilution
Lane 1:
Wild-type Neuro-2a cell lysate at 20 µg
Lane 2:
ARC knockout Neuro-2a cell lysate at 20 µg
Lane 2:
Western blot - Mouse ARC knockout Neuro-2a cell line (ab280071)
Lane 3:
SH-SY5Y cell lysate at 20 µg
Lane 4:
K562 cell lysate at 20 µg
Lane 5:
Mouse Brain cell lysate at 20 µg
Predicted band size: 45 kDa
Observed band size: 45 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Mouse ARC knockout Neuro-2a cell line (AB280071)
Complementary Products
Primary Antibodies
AB183183
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Arc antibody [EPR18950] (AB183183)](https://content.abcam.com/products/images/arc-antibody-epr18950-ab183183--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img42591.jpg)
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Primary Antibodies
AB76026
Anti-Nrf2 (phospho S40) antibody [EP1809Y]
20ul selling size|RabMAb|Recombinant
![Western blot - Anti-Nrf2 (phospho S40) antibody [EP1809Y] (AB76026)](https://content.abcam.com/products/images/nrf2-phospho-s40-antibody-ep1809y-ab76026--western-blot-img21503.jpg)
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Primary Antibodies
AB108349
Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal
BOND RX™ Validated|20ul selling size|KO Validated|RabMAb|Recombinant
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CDKN2A/p16INK4a antibody [EPR1473] - C-terminal (AB108349)](https://content.abcam.com/products/images/cdkn2a-p16ink4a-antibody-epr1473-c-terminal-ab108349--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img74532.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 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 for bath 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 - 5x105 cells/mL(for initial passages it is recomended to culture the cells in the higher range of recomended seeding density). 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.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x105 cells/mL is recommended.
Culture medium
EMEM + 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
The Arc protein influences synaptic plasticity and memory formation. It does not operate alone; instead it interacts within complexes that regulate synaptic strength. These complexes contain other proteins critical for actin cytoskeleton remodeling which impacts synaptic efficacy. Through these interactions Arc supports the internalization of AMPA receptors playing a role in long-term depression (LTD) and long-term potentiation (LTP) of synapses essential mechanisms for learning and memory.
Pathways
The Arc protein forms integral parts of the synaptic signaling pathways. It connects with the BDNF-TrkB signaling pathway which is important for neuronal development and synaptic modulation. Arc also relates to the mTOR signaling pathway which governs cellular growth and protein synthesis. Through these pathways Arc interacts with proteins like Neuro2a and SynGAP highlighting its comprehensive role in synaptic regulation and plasticity.
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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