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- 抗原 See all SUN5 抗体
- SUN5 (Sad1 and UNC84 Domain Containing 5 (SUN5))
- 适用
- 人, 小鼠, 大鼠
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宿主
- 兔
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克隆类型
- 多克隆
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标记
- This SUN5 antibody is un-conjugated
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应用范围
- Western Blotting (WB)
- 产品特性
- Expected molecular weight of target: 379 kDa
- 纯化方法
- Affinity purification
- 亚型
- IgG
- Top Product
- Discover our top product SUN5 Primary Antibody
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- 应用备注
- WB 1:500-1:2000
- 限制
- 仅限研究用
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- by
- Johann-Friedrich-Blumenbach-Institute for Zoology and Anthropology, Department of Developmental Biology, Georg-August-University Göttingen
- No.
- #102344
- 日期
- 2017.12.19
- 抗原
- SUN5
- Lot Number
- Method validated
- Immunocytochemistry
- Positive Control
- NIH/3T3 mouse embryonic fibroblast cells overexpressing HA- and myc-tagged SPAG4L-2/SUN5
- Negative Control
- NIH/3T3 mouse embryonic fibroblast cells overexpressing HA- and myc-tagged SPAG4
- Notes
Passed. ABIN1876373 is specific for SPAG4L-2/SUN5 and did not detect SPAG4 when ectopically expressed. Unspecific binding to other cellular proteins is nearly absent.
- Primary Antibody
- ABIN1876373
- Secondary Antibody
- goat anti-rabbit IgG DyLight488 (Thermo Scientific, 35553, lot KA130088)
- Full Protocol
- NIH/3T3 cells (ATCC, CRL-1658) are grown in DMEM+GlutaMAX (Gibco, 31966-021, Lot 1852045) supplemented with fetal bovine serum (Gibco 270-106) and Pen/Strep (Gibco 15140), at 37°C and 5% CO2 to 70% confluency.
- Transfect cells with HA/myc-tagged SPAG4 expression plasmid or HA/myc-tagged SPAG4L-2/SUN5 (Frohnert et al., 2011) expression plasmid using EndofectinMax (GeneCopoeia) following the manufacturer's instructions.
- Grow cells for 24h.
- Fix cells in 3.7% paraformaldehyde (in PBS) for 15min at 4°C followed by incubation in 0.3% Triton X-100 for 10min at 4°C.
- Block cells in PBS containing 1% bovine serum albumin and 0.5% Tween-20 (PBT) for 1h at room temperature.
- Incubate cells with primary
- rabbit anti-SUN5 antibody (antibodies-online, ABIN1876373) diluted 1:100 in PBS ON at 4° and
- mouse anti-myc tag antibody clone (9E10) diluted 1:100 in PBS ON at 4°C.
- Wash cells with TBST (50mM Tris-HCl, pH 7.4, 150mM NaCl, 0.1% Tween 20) for 15min at RT.
- Incubate cells with secondary
- goat anti-rabbit IgG DyLight488 (Thermo Scientific, 35553, lot KA130088) diluted 1:1000 in PBS for 1h at 37°C and
- goat anti-mouse IgG (H+L) Alexa Fluor 555 (Invitrogen, A21422, lot 948498) diluted 1:1000 in PBS for 1h at 37°C.
- Counterstain DNA with DAPI (4,6-Diamidino-2-phenylindole; Sigma D-9542).
- Image acquisition on Zeiss LSM 510 confocal microscope and processing using Adobe Photoshop 5.0.
- Experimental Notes
生效 #102344 (Immunocytochemistry)Validation ImagesSpecificity of ABIN1876373. Cells were transfected either with SPAG4 (upper row) or SPAG4L-2/SUN5 (lower row), both tagged with HA and myc. Anti-myc antibody (B, F; red) detected both proteins, but ABIN1876373 (A, E; green) exclusively detected SPAG4L-2/SUN5 without any cross-reaction to SPAG4. The antibody detected SPAG4L-2/SUN5 at the nuclear membrane, as expected. The pictures on the right (D, H) show the red and green channels merged with DAPI counterstain (blue).Full Methods -
- 状态
- Liquid
- 浓度
- 1 mg/mL
- 缓冲液
- PBS with 0.02 % sodium azide, 50 % glycerol, pH 7.4
- 储存液
- Sodium azide
- 注意事项
- This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- 注意事项
- Avoid repeated freeze/thaw cycles.
- 储存条件
- -20 °C
- 储存方法
- Store at -20°C.
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- 抗原
- SUN5 (Sad1 and UNC84 Domain Containing 5 (SUN5))
- 别名
- SPAG4L (SUN5 产品)
- 别名
- SPAG4L antibody, TSARG4 antibody, dJ726C3.1 antibody, 1700021O15Rik antibody, SPAG4L-2 antibody, SRG4 antibody, Spag4l antibody, Sad1 and UNC84 domain containing 5 antibody, SUN5 antibody, Sun5 antibody
- 基因ID
- 140732
- UniProt
- Q8TC36
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