- Sanger seq
Supplier Data
Sanger Sequencing - Human RAB3A knockout SK-N-FI cell line (AB288708)
Homozygote. 62 bp deletion
- WB
Lab
Western blot - Human RAB3A knockout SK-N-FI cell line (AB288708)
ab302518 was shown to react with RAB3A in wild-type SKNFI cells in Western blot with loss of signal observed in a RAB3A knockout cell line. Wild-type SKNFI and RAB3A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab302518 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-Rab3A antibody [EPR26022-9] (<a href='/ko/products/primary-antibodies/rab3a-antibody-epr26022-9-ab302518'>ab302518</a>) at 1/1000 dilution
Lane 1:
Wild-type SKNFI lysate at 40 µg
Lane 2:
Western blot - Human RAB3A knockout SK-N-FI cell line (ab288708) at 40 µg
false
- WB
Lab
Western blot - Human RAB3A knockout SK-N-FI cell line (AB288708)
Anti-RAB3A antibody [EPR26022-9] (ab302518) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab302518 was shown to bind specifically to RAB3A. A band was observed at 27 kDa in wild-type SKNFI cell lysates with no signal observed at this size in RAB3A knockout cell line. To generate this image, wild-type and RAB3A knockout SKNFI cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% 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-Rab3A antibody [EPR26022-9] (<a href='/ko/products/primary-antibodies/rab3a-antibody-epr26022-9-ab302518'>ab302518</a>) at 1/1000 dilution
Lane 1:
Wild-type SKNFI cell lysate at 20 µg
Lane 2:
Western blot - Human RAB3A knockout SK-N-FI cell line (ab288708) at 20 µg
Lane 3:
Human brain cell lysate at 20 µg
Lane 4:
HEK-293 cell lysate at 20 µg
Observed band size: 27 kDa
false
Reactivity 정보
제품 세부 정보
Recommended control: Human wild-type SK-N-FI cell line ( ab288716). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
Cryopreservation cell medium: Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture medium: DMEM + 10% FBS
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.
- 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 2x104cells/cm2. 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 2x104 cells/cm2 is recommended.
- A partial media change 24 hours prior to subculture may be helpful to encourage growth, if required.
- Cells should be passaged when they have achieved 80-90% confluence.
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.
특성 및 보관 정보
유전자명
유전자 편집 종류
유전자 편집 방식
Knockout 검증
배송 시 보관 조건
적절한 단기 보관 기간
적절한 단기 보관 조건
적절한 장기 보관 조건
추가 정보
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Rab3A facilitates neurotransmitter release by interacting with effector proteins such as RIM and Rabphilin-3A. It is part of a larger complex that includes the SNAREs which help mediate synaptic vesicle fusion. Through its regulatory action Rab3A ensures that neurotransmitters are released in a controlled manner during synaptic transmission which is essential for proper neuronal communication.
Pathways
Rab3A participates in the synaptic vesicle cycle which is vital in neurotransmitter release. This cycle involves key proteins like Syntaxin and VAMP. Rab3A interacts with these SNARE proteins to participate in vesicle docking and priming. Additionally it plays a role in the calcium-dependent exocytosis pathway which relies on calcium ions triggering vesicle fusion at nerve terminals.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
초기 취급 지침
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 가이드라인
- 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.
배양 배지
DMEM + 10% FBS
동결보존 배지
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
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