SUN5 抗体
(1 validation)Quick Overview for SUN5 抗体 (ABIN1876373)
抗原
See all SUN5 抗体适用
宿主
克隆类型
标记
应用范围
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产品特性
- Expected molecular weight of target: 379 kDa
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纯化方法
- Affinity purification
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亚型
- IgG
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anti-Sad1 and UNC84 Domain Containing 5 (SUN5) (AA 161-275) antibody
SUN5 适用: 小鼠, 大鼠 WB, ELISA, IF (cc), IF (p), IHC (fro), IHC (p) 宿主: 兔 Polyclonal unconjugated
anti-Sad1 and UNC84 Domain Containing 5 (SUN5) (Internal Region) antibodySUN5 适用: 人 ELISA, IHC 宿主: 兔 Polyclonal unconjugated
anti-Sad1 and UNC84 Domain Containing 5 (SUN5) (AA 1-379) antibodySUN5 适用: 人 WB 宿主: 小鼠 Polyclonal unconjugated
anti-Sad1 and UNC84 Domain Containing 5 (SUN5) (N-Term) antibodySUN5 适用: 人 WB 宿主: 兔 Polyclonal unconjugated
anti-Sad1 and UNC84 Domain Containing 5 (SUN5) (N-Term) antibodySUN5 适用: 人 WB 宿主: 兔 Polyclonal unconjugated
anti-Sad1 and UNC84 Domain Containing 5 (SUN5) (N-Term) antibodySUN5 适用: 人 WB 宿主: 兔 Polyclonal unconjugated
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应用备注
- WB 1:500-1:2000
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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)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![Specificity 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).]()
Specificity 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 -
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状态
- Liquid
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浓度
- 1 mg/mL
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缓冲液
- PBS with 0.02 % sodium azide, 50 % glycerol, pH 7.4
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储存液
- Sodium azide
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注意事项
- This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
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注意事项
- Avoid repeated freeze/thaw cycles.
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储存条件
- -20 °C
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储存方法
- Store at -20°C.
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- SUN5 (Sad1 and UNC84 Domain Containing 5 (SUN5))
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别名
- SPAG4L
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基因ID
- 140732
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UniProt
- Q8TC36
抗原
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