RND3 抗体 (Center)
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- 抗原 See all RND3 抗体
- RND3 (rho Family GTPase 3 (RND3))
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抗原表位
- Center
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适用
- 人, 小鼠
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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 RND3 antibody is un-conjugated
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应用范围
- Western Blotting (WB), ELISA, Immunohistochemistry (IHC)
- 预测反应
- Human Mouse
- 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.
- 亚型
- Ig Fraction
- Top Product
- Discover our top product RND3 Primary Antibody
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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
- 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)Validation ImagesFull Methods -
- 状态
- Liquid
- 储存条件
- 4 °C,-20 °C
- 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.
- 有效期
- 6 months
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- 抗原
- RND3 (rho Family GTPase 3 (RND3))
- 别名
- RND3 (RND3 产品)
- 别名
- ARHE antibody, Rho8 antibody, RhoE antibody, memB antibody, MGC83857 antibody, 2610017M01Rik antibody, AI661404 antibody, Arhe antibody, Rhoe antibody, RHOE antibody, arhe antibody, id:ibd2712 antibody, id:ibd2919 antibody, rho8 antibody, rnd3 antibody, wu:fc39h07 antibody, zgc:73080 antibody, wu:fd06e12 antibody, zgc:92324 antibody, Rho family GTPase 3 antibody, Rho family GTPase 3 L homeolog antibody, Rho family GTPase 3a antibody, Rho family GTPase 3b antibody, RND3 antibody, rnd3.L antibody, Rnd3 antibody, rnd3a antibody, rnd3b antibody
- 分子量
- 27368 Da
- 基因ID
- 390
- NCBI登录号
- NP_001241667
- UniProt
- P61587
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