SPNS2 抗体 (N-Term)
(1 validation)-
- 抗原 See all SPNS2 抗体
- SPNS2 (Spinster Homolog 2 (SPNS2))
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抗原表位
- N-Term
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适用
- 人, 小鼠, 大鼠, Pig, 兔
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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 SPNS2 antibody is un-conjugated
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应用范围
- Western Blotting (WB)
- 序列
- PPGTPGTPGC AATAKGPGAQ QPKPASLGRG RGAAAAILSL GNVLNYLDRY
- 预测反应
- Human: 100%, Mouse: 85%, Pig: 100%, Rabbit: 92%, Rat: 93%
- 产品特性
- This is a rabbit polyclonal antibody against SPNS2. It was validated on Western Blot using a cell lysate as a positive control.
- 纯化方法
- Affinity Purified
- 免疫原
- The immunogen is a synthetic peptide directed towards the N terminal region of human SPNS2
- Top Product
- Discover our top product SPNS2 Primary Antibody
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anti-Spinster Homolog 2 (SPNS2) (AA 68-94), (N-Term) antibody
SPNS2 适用: 人 WB, IHC (p), FACS 宿主: 兔 Polyclonal RB38003 unconjugated
anti-Spinster Homolog 2 (SPNS2) (AA 71-120) antibodySPNS2 适用: 人, 小鼠, 猴, Pig, Bat WB, IHC, IHC (p) 宿主: 兔 Polyclonal unconjugated
anti-Spinster Homolog 2 (SPNS2) (AA 1-140) antibodySPNS2 适用: 人 ELISA, IF 宿主: 兔 Polyclonal unconjugated
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- 应用备注
- Optimal working dilutions should be determined experimentally by the investigator.
- 说明
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Antigen size: 549 AA
- 限制
- 仅限研究用
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- by
- Puissant lab, Institute of Hematology, Inserm U944, CNRS UMR 7212
- No.
- #101829
- 日期
- 2018.02.01
- 抗原
- SPNS2
- Lot Number
- QC66273-42972
- Method validated
- Western Blotting
- Positive Control
- Baf-3 cells/ MLL-AF9 murine primary cell model
- Negative Control
- Baf-3 or MLL-AF9 murine primary cell model silenced for SPNS2 gene with 2 different shRNA
- Notes
Passed. ABIN2786494 specifically recognizes SPNS2 in murine Baf-3 cells or MLL-AF9 primary cell model.
- Primary Antibody
- ABIN2786494
- Secondary Antibody
- mouse anti-rabbit HRP conjugated antibody (GE Healthcare, NA9340V)
- Full Protocol
- Grow Baf-3 or MLL-AF9 cells in RPMI 1640 1x (Gibco, 21875-059) supplemented with 10% FBS (Sigma-Aldrich, F6178, lot 16G148) and 1x Penicillin/Streptomycin (Gibco, 15140-122), at 37°C and 5% CO2 in 10ml on a flask to 70% confluency.
- Transduce 3x106 cells with 35µl of concentrated retrovirus, spin-infection (3h at 37°C to 1200 rpm).
- Grow cells for 3d.
- Induce and select cells.
- Grow cells for 5d.
- Sort and collect cells (doxycycline).
- Lyse 7x105 cells in 25µl per well in cold SDS Lysis Buffer (Cell Signaling).
- Determine total protein content of the lysates using Bradford assay (Bio-Rad, 5000006).
- Denature 20µg of total protein for 7min at 95°C in 6µl Laemmli SDS sample buffer and subsequently separate them on a denaturing NuPAGE 4-12% Bis-Tris Protein Gels (Invitrogen,NP0321BOX ) in an ELECTROPHORESIS CHAMBER for 2h at 150V.
- Transfer proteins onto PVDF membrane (Millipore) with a Western blotting system for 1h 40min at 300mA.
- Block the membrane with blocking buffer (2.75% gelatine, 30% BSA, 10mM Tris, NaCl, EDTA, tween 20) for 1h at RT.
- Incubation with rabbit anti-SPNS2 antibody (antibodies-online, ABIN2786494, lot QC66273-42972) diluted 1:500 in blocking buffer for 16h at 4°C.
- Wash membrane 3x for 10min with tween TBS buffer.
- Incubation with secondary mouse anti-rabbit HRP conjugated antibody (GE Healthcare, NA9340V) diluted 1:5000 in blocking buffer for 1h at RT.
- Wash membrane 3x for 10min with TBST buffer.
- Reveal protein bands using Pierce ECL Western Blotting Substrate (ThermoFisher Scientific, 32106) on a Li-Cor imaging machine.
- Experimental Notes
The SPNS2 antibody ABIN2786494 reveals a protein of the expected molecular weight for SPNS2 in lysates of murine Baf-3 cells or MLL-AF9 primary cell model.
ABIN2786494 does show some unspecific binding but the SPNS2 band itself is clear and only visible in the positive but not the negative controls.
生效 #101829 (Western Blotting)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![Western blot analysis of SPNS2 expression in mouse Baf-3 (left) and MLL-AF9 cells (right) using ABIN2786494. Cells were transduced either with a control shRNA (control shRNA) or one of two SPNS2 shRNA (shSPNS2_1 and shSPNS2_5).]()
Western blot analysis of SPNS2 expression in mouse Baf-3 (left) and MLL-AF9 cells (right) using ABIN2786494. Cells were transduced either with a control shRNA (control shRNA) or one of two SPNS2 shRNA (shSPNS2_1 and shSPNS2_5).
Full Methods -
- 状态
- Liquid
- 浓度
- Lot specific
- 缓冲液
- Liquid. Purified antibody supplied in 1x PBS buffer with 0.09 % (w/v) sodium azide and 2 % sucrose.
- 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
- For short term use, store at 2-8°C up to 1 week. For long term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.
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- 抗原
- SPNS2 (Spinster Homolog 2 (SPNS2))
- 别名
- SPNS2 (SPNS2 产品)
- 别名
- fi20h04 antibody, si:dkey-7b17.4 antibody, toh antibody, wu:fi20h04 antibody, DKFZp459J1933 antibody, spinster homolog 2 (Drosophila) antibody, sphingolipid transporter 2 antibody, spinster homolog 2 antibody, SPNS2 antibody, spns2 antibody, Spns2 antibody
- 背景
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SPNS2 is the sphingolipid transporter required for migration of myocardial precursors. SPNS2 transports sphingosine 1-phosphate (S1P), a secreted lipid mediator that plays critical roles in cardiovascular, immunological, and neural development and function. SPNS2 mediates the export of S1P from cells in the extraembryonic yolk syncytial layer (YSL), thereby regulating myocardial precursor migration.
Alias Symbols: -
Protein Size: 549 - 分子量
- 60 kDa
- 基因ID
- 124976
- NCBI登录号
- NM_001124758, NP_001118230
- UniProt
- Q8IVW8
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