RND3 抗体 (Center)
(1 validation)Quick Overview for RND3 抗体 (Center) (ABIN2496515)
抗原
See all RND3 抗体适用
宿主
克隆类型
标记
应用范围
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抗原表位
- Center
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预测反应
- Human Mouse
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免疫原
- This ARHE antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 142~171 amino acids from the Center region of human AEHE.
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亚型
- Ig Fraction
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anti-rho Family GTPase 3 (RND3) (AA 104-241) antibody
RND3 适用: 人 ELISA, IHC, ICC, FACS 宿主: 小鼠 Monoclonal 5C7E8 unconjugated
anti-rho Family GTPase 3 (RND3) (AA 1-244) antibodyRND3 适用: 人 ELISA, WB, IHC (p) 宿主: 小鼠 Monoclonal 1D2 unconjugated
anti-rho Family GTPase 3 (RND3) (AA 1-245) antibodyRND3 适用: 人 ELISA, WB, IHC, IHC (p) 宿主: 小鼠 Monoclonal 1D2 unconjugated
anti-rho Family GTPase 3 (RND3) (Internal Region) antibodyRND3 适用: 人, 小鼠, 大鼠 WB, IHC, ICC, IF 宿主: 兔 Polyclonal unconjugated
anti-rho Family GTPase 3 (RND3) (AA 133-165) antibodyRND3 适用: 人, 小鼠, 大鼠 WB, IHC (p) 宿主: 兔 Polyclonal RB14500 unconjugated
anti-rho Family GTPase 3 (RND3) (AA 152-239) antibodyRND3 适用: 人 ELISA, IHC 宿主: 兔 Polyclonal unconjugated
anti-rho Family GTPase 3 (RND3) (AA 1-244) antibodyRND3 适用: 人 WB 宿主: 小鼠 Polyclonal unconjugated
anti-rho Family GTPase 3 (RND3) (C-Term) antibodyRND3 适用: 人, 小鼠, 大鼠, Pig, Cow, 豚鼠, 马, 兔 WB 宿主: 兔 Polyclonal unconjugated
anti-rho Family GTPase 3 (RND3) (AA 1-245) antibodyRND3 适用: 人 WB, IHC (p), EIA 宿主: 小鼠 Monoclonal 1D2 unconjugated
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应用备注
- Optimal working dilution should be determined by the investigator.
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限制
- 仅限研究用
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- by
- Instituto de Parasitología y Biomedicina López-Neyra
- No.
- #100103
- 日期
- 2016.12.23
- 抗原
- RND3
- Lot Number
- SH080408I
- Method validated
- Western Blotting
- Positive Control
- primary mouse microglia
- Negative Control
- siRNA knock-down of RND3 in primary mouse microglia
- Notes
- Passed. ABIN2496515 specifically recognizes Rnd3 in primary mouse microglia lysates.
- Primary Antibody
- ABIN2496515
- Secondary Antibody
- anti-rabbit HRP-conjugated antibodies (DakoCytomation, P0448, lot 00055814)
- Full Protocol
- Primary mouse microglia are grown in DMEM (Invitrogen), supplemented with 10% foetal bovine serum (Gibco), 10% horse serum and penicillin and streptomycin (Gibco), at 37°C and 5% CO2 in a 12-well dish.
- Transfect cells with cells with 0.05nmol mouse RND3 Silencer Select siRNA (ThermoFisher Scientific, ASO10960, ASO109AE and ASO109FC, lot AMO04BXD) using Lipofectamine 3000 (Invitrogen) following the manufacturer´s instructions.
- Lyse cells in 25µl per well cold lysis buffer (10mM Tris-HCl pH 8.0, 150mM NaCl, 1% Nonidet-P40, 1mM EDTA, 10mM NaF, 1mM Na3VO4, protease inhibitors (Sigma)).
- Determine total protein content of the lysates using Bradford assay (Bio-Rad, 500-0006, lot 111832).
- Denature 5µg total protein for 5min at 95°C in 5µl Laemmli SDS sample buffer and subsequently separate them on a denaturing freshly cast 10% SDS-PAGE for 1h at 140V.
- Transfer proteins onto PVDF membrane (Pall Life Sciences, 75696G, lot TO3225) with a semi-dry Western blotting system for 50min at 60mA.
- Block the membrane with TBST (TBS, 0.1% Tween) containing 5% milk ON at 4°C.
- Incubation with primary mouse anti-RND3 antibody (antibodies-online, ABIN2496515, lot SH080408I) diluted 1:500 in TBST containing 5% milk for 1h at RT.
- Wash membrane 5x 5min with TBST.
- Incubation with anti-rabbit horseradish peroxidase-conjugated secondary antibodies (DakoCytomation, P0448, lot 00055814) diluted 1:2000 in TBST containing 5% milk for 1h at RT.
- Wash membrane 5x 5min with TBST and 3x 5min with TBS.
- Reveal protein bands using ECL Prime Western Blotting Detection Reagent (GE Healthcare, RPN2232, lot 9547481) with Curix RP2 Plus medical X-ray films (AGFA Health Care, ENKMV, lot 79040074) on an AGFA Health Care Curix60-developer.
- Strip membranes using RestoreTM Western Blot Stripping Buffer (Thermo Scientific, 21059, lot NA 164269) and subsequently incubate with a GAPDH loading control antibody (Sigma, G9545, lot 049K4787) diluted 1:10000 and with anti-rabbit horseradish peroxidase-conjugated secondary antibodies (DakoCytomation, P0448, lot 00055814) diluted 1:2000.
- Wash and reveal the protein bands using self-made ECL Western Blotting Detection Reagent with Curix RP2 Plus medical X-ray films (AGFA Health Care, ENKMV, lot 79040074) on an AGFA Health Care Curix60-developer.
- Experimental Notes
- The RND3 antibody ABIN2496515 reveals a protein of the expected molecular weight of murine Rnd3 in lysates of primary mouse microglia. The protein band’s intensity decreases upon siRNA knockdown.
生效 #100103 (Western Blotting)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![Primary mouse microglia were transfected with Rnd3 siRNA and Rnd3 was revealed using ABIN2496515 as described in the protocol section (lanes 1, 2, and 3). Lysates from cells transfected with scrambled siRNA (lane 4) and untransfected cells (lane 5) served as negative controls.]()
Primary mouse microglia were transfected with Rnd3 siRNA and Rnd3 was revealed using ABIN2496515 as described in the protocol section (lanes 1, 2, and 3). Lysates from cells transfected with scrambled siRNA (lane 4) and untransfected cells (lane 5) served as negative controls.
Full Methods -
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状态
- Liquid
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储存条件
- 4 °C,-20 °C
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储存方法
- Maintain refrigerated at 2-8 °C for up to 6 months. For long term storage store at -20 °C in small aliquots to prevent freeze-thaw cycles.
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有效期
- 6 months
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- RND3 (rho Family GTPase 3 (RND3))
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别名
- RND3
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分子量
- 27368 Da
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基因ID
- 390
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NCBI登录号
- NP_001241667
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UniProt
- P61587
抗原
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