SRSF5 抗体 (AA 1-272)
(1 validation)-
- 抗原 See all SRSF5 抗体
- SRSF5 (serine/arginine-Rich Splicing Factor 5 (SRSF5))
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抗原表位
- AA 1-272
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适用
- 人
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宿主
- 小鼠
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克隆类型
- 多克隆
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标记
- This SRSF5 antibody is un-conjugated
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应用范围
- Western Blotting (WB), Immunofluorescence (IF)
- 原理
- Mouse polyclonal antibody raised against a full-length human SFRS5 protein.
- 序列
- MSGCRVFIGR LNPAAREKDV ERFFKGYGRI RDIDLKRGFG FVEFEDPRDA DDAVYELDGK ELCSERVTIE HARARSRGGR GRGRYSDRFS SRRPRNDRRN APPVRTENRL IVENLSSRVS WQDLKDFMRQ AGEVTFADAH RPKLNEGVVE FASYGDLKNA IEKLSGKEIN GRKIKLIEGS KRHSRSRSRS RSRTRSSSRS RSRSRSRSRK SYSRSRSRSR SRSRSKSRSV SRSPVPEKSQ KRGSSSRSKS PASVDRQRSR SRSRSRSVDS GN
- 交叉反应
- 人
- 产品特性
- Antibody reactive against mammalian transfected lysate.
- 免疫原
- SFRS5 (NP_001034554.1, 1 a.a. ~ 272 a.a) full-length human protein.
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anti-serine/arginine-Rich Splicing Factor 5 (SRSF5) (AA 1-272) antibody
SRSF5 适用: 人 WB, IF, IHC (p) 宿主: 小鼠 Polyclonal unconjugated
anti-serine/arginine-Rich Splicing Factor 5 (SRSF5) (AA 1-272) antibodySRSF5 适用: 人 ELISA, WB 宿主: 小鼠 Monoclonal 2C8 unconjugated
anti-serine/arginine-Rich Splicing Factor 5 (SRSF5) antibodySRSF5 适用: 人, 小鼠, 大鼠, 猴 WB, IF, IHC, IC 宿主: 兔 Polyclonal unconjugated
anti-serine/arginine-Rich Splicing Factor 5 (SRSF5) (AA 70-97) antibodySRSF5 适用: 人 WB, FACS 宿主: 兔 Polyclonal RB19850 unconjugated
anti-serine/arginine-Rich Splicing Factor 5 (SRSF5) (AA 71-120) antibodySRSF5 适用: 人, 小鼠, 大鼠 ELISA, IF 宿主: 兔 Polyclonal unconjugated
anti-serine/arginine-Rich Splicing Factor 5 (SRSF5) (Middle Region) antibodySRSF5 适用: 人 WB, EIA 宿主: 兔 Polyclonal unconjugated
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- 应用备注
- Optimal working dilution should be determined by the investigator.
- 限制
- 仅限研究用
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- by
- Laboratoire de Biologie et Modélisation de la Cellule, ENS Lyon
- No.
- #102043
- 日期
- 2017.11.27
- 抗原
- SRSF5
- Lot Number
- 11159
- Method validated
- Western Blotting
- Positive Control
- MDA-MB-231 control siRNA, MCF-7 control siRNA
- Negative Control
- qPCR verified SRSF5 siRNA knockdown in MDA-MB-231 and MCF-7 cells
- Notes
- ABIN948457 specifically reveals SRSF5 in human MDA-MB-231 and MCF-7 cell extracts but also several extraneous protein bands.
- Primary Antibody
- ABIN948457
- Secondary Antibody
- HRP-conjugated donkey anti-mouse antibody (Bethyl Laboratories, A90-137P)
- Full Protocol
- Grow cells in DMEM (MCF7: Gibco, 10938-025, lot 185560; MDA-MB-231: Gibco, 11880-028, lot 1894718) supplemented with FBS (Gibco, 131608-01, lot 42F6573K) and penicillin-streptomycin (Gibco, 15140-122, lot 1902417), and L-glutamine (Gibco, 25030-024, lot 1895805) 37°C and 5% CO2.
- Transfect 2.5x106 cells with amount 50nmol of siRNA (eurofins genomics) using Lipofectamine RNAiMAX reagent (Invitrogen, 13778030, lot 1863033) following the manufacturer´s instructions.
- Grow cell for 48h.
- Lyse 5x106 cells in 100µl of cold lysis buffer (50mM Tris-HCl, 400mM NaCl, 5mM EDTA pH8.0, 1% NP40, 0.2% SDS) containing 1x protease inhibitor cocktail (Roche, 05056489001).
- Determine total protein content of the lysates using the Pierce BCA protein assay (ThermoFisher Scientific, 23225, lot sc246917).
- Denature 30µg of total protein for 5min at 95°C in 30µl using NuPAGE Sample Reducing reagent 10x (Invitrogen, np0009, lot 1879834) and the LDS sample buffer (4X) (ThermoFisher Scientific, np0007, lot 1879570) and subsequently separate them on a denaturing gel Bis-Tris 4-12% Protein Gels (ThermoFisher Scientific, np0336box, lot 17061471) in MOPS migration buffer (ThermoFisher Scientific, NP0001, lot 1881690) for 1h at 160V.
- Transfer proteins onto a Trans-Blot Turbo Mini Nitrocellulose membrane (Bio-Rad, 170-4158, lot 64126951) with a Transblot Turbo Transfer System (Bio-Rad, 1704150) for 7min at 1.3A and 25V.
- Block the membrane with TBST (50mM Tris-HCl pH7.4-7.6, 150mM NaCl, 0.1% Tween 20) containing 5% non-fat milk for 1h with slight agitation at RT.
- Incubation with primary mouse anti-human SRSF5 antibody (antibodies-online, ABIN948457, lot 11159) diluted to 1µg/µl in 5ml final volume in TBST + milk ON at 4°C.
- Wash membrane 2x for 10min with TBST.
- Incubation with secondary HRP-conjugated donkey anti-mouse antibody (Bethyl Laboratories, A90-137P) diluted 1:10000 in TBST + milk for 1h at RT.
- Wash membrane 2x for 10min with TBST.
- Reveal protein bands using ECL Prime Western Blotting Detection Reagent (GE healthcare RPN2236, lot 9794767) on a ChemiDoc MP Imaging System (Bio-Rad).
- Experimental Notes
SRSF5 was depleted to almost 70% in both tested cell lines (Figure A). Quantities are normalized relative to the control siRNA (siGL2) and in comparison with a housekeeping gene (ribosomal 18S RNA, not shown). SRSF5 primers were previously validated on both cell lines. Efficacity of the primers are for the calculation estimated at 2.
ABIN948457 specifically reveals a protein band with an apparent MW of approximately 45kDa in cell lysates which were transfected with control siRNA (Figure B). Post-translational modifications such as phosphorylation can influence the migration of SRSF5, making it appear bigger than it really is. The band disappears upon knockdown of the SRSF5 expression in cells transfected with an siSRSF5 construct. The antibody does also detect several other proteins at approximately 35-38kDa and 55kDa.
生效 #102043 (Western Blotting)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![A RT-qPCR verified depletion of SRSF5 transcription in MDA-MB-231 (MDA) and MCF-7 cells (MCF) subsequently to transfection with SRSF5 siRNA (siSRSF5) or control siRNA (siGL2). B Western blot analysis using ABIN948457 on cellular extracts with or without siRNA SRSF5 knock-down.]()
A RT-qPCR verified depletion of SRSF5 transcription in MDA-MB-231 (MDA) and MCF-7 cells (MCF) subsequently to transfection with SRSF5 siRNA (siSRSF5) or control siRNA (siGL2). B Western blot analysis using ABIN948457 on cellular extracts with or without siRNA SRSF5 knock-down.
Full Methods -
- 缓冲液
- In 1x PBS, pH 7.4
- 注意事项
- Aliquot to avoid repeated freezing and thawing.
- 储存条件
- -20 °C
- 储存方法
- Store at -20°C or lower. Aliquot to avoid repeated freezing and thawing.
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- 抗原
- SRSF5 (serine/arginine-Rich Splicing Factor 5 (SRSF5))
- 别名
- SFRS5 (SRSF5 产品)
- 别名
- HRS antibody, SFRS5 antibody, SRP40 antibody, Sfrs5 antibody, SRp40 antibody, hrs antibody, sfrs5 antibody, srp40 antibody, srsf5 antibody, sfrs5b antibody, wu:fe15e05 antibody, zgc:113907 antibody, zgc:92278 antibody, serine and arginine rich splicing factor 5 antibody, serine/arginine-rich splicing factor 5 antibody, serine/arginine-rich splicing factor 5 S homeolog antibody, serine/arginine-rich splicing factor 5b antibody, SRSF5 antibody, Srsf5 antibody, srsf5.S antibody, srsf5b antibody
- 背景
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Full Gene Name: splicing factor, arginine/serine-rich 5
Synonyms: HRS,SRP40 - 基因ID
- 6430
- NCBI登录号
- NM_001039465
- 途径
- Ribonucleoprotein Complex Subunit Organization
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