RHBDF2 抗体 (N-Term)
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北京 101111
Quick Overview for RHBDF2 抗体 (N-Term) (ABIN2846535)
抗原
See all RHBDF2 抗体适用
宿主
克隆类型
标记
应用范围
克隆位点
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抗原表位
- AA 80-109, N-Term
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纯化方法
- This antibody is purified through a protein A column, followed by peptide affinity purification.
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免疫原
- This RHBDF2 antibody is generated from rabbits immunized with a KLH conjugated synthetic peptide between 80-109 amino acids from the N-terminal region of human RHBDF2.
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亚型
- IgG
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anti-Rhomboid 5 Homolog 2 (RHBDF2) (AA 80-109), (N-Term) antibody
RHBDF2 适用: 人, 小鼠 WB, IHC (p) 宿主: 兔 Polyclonal RB32834 unconjugated
anti-Rhomboid 5 Homolog 2 (RHBDF2) (AA 1-200) antibodyRHBDF2 适用: 人 ELISA, IHC 宿主: 兔 Polyclonal unconjugated
anti-Rhomboid 5 Homolog 2 (RHBDF2) (AA 1-200) antibodyRHBDF2 适用: 人 ELISA, IHC, IF 宿主: 兔 Polyclonal unconjugated
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应用备注
- WB: 1:2000. WB: 1:2000. WB: 1:1000. IHC-P: 1:25. IHC-P: 1:25
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限制
- 仅限研究用
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- by
- Prof. Ludwig
- No.
- #104634
- 日期
- 2026.01.19
- 抗原
- RHBDF2
- Lot Number
- SA180724YE
- Method validated
- Western Blotting
- Positive Control
Overexpressed human and murine iRhom2.
- Negative Control
HEK293 iRhom1/iRhom2 double knockout.
- Notes
The iRhom2 antibody ABIN2846535 works in WB at a 1:1000 dilution to detect overexpressed human iRhom2. Non-specific bands are also detected.
- Primary Antibody
- ABIN2846535
- Secondary Antibody
- Full Protocol
- Lyse HEK293 cells with lysis buffer (50 mM Tris, 137 mM NaCl, 2 mM EDTA, and 10 mM 1,10-Phenanthroline, adjusted to pH 7.5 and supplemented with 1% Triton X-100 (Sigma-Aldrich) and cOmplete™ protease inhibitor cocktail (Sigma-Aldrich)) for 40 min at 4°C.
- Clear lysates by centrifugation at 20,000 rpm for 20 min at 4°C.
- Denature samples with reducing Lämmli buffer (3% (w/v) SDS, 16% glycerol, 8% 2-mercaptoethanol, 0.01% (w/v) bromophenol blue, and 0.1 M Tris-HCl at pH 6.8) for 20 min at 60°C.
- Separate protein samples on a denaturing 12% SDS-PAGE gel alongside a PageRuler™ Plus Prestained Protein Ladder (Thermo Fisher, 26619).
- Wet the Immobilon®-FL PVDF membrane (0.45 µm pore size) (Millipore, IPFL00010) in methanol for 30 sec.
- Wash membrane once with Milli-Q® water for 1 min.
- Electro-transfer proteins onto Immobilon®-FL PVDF membrane (0.45 µm pore size) (Millipore, IPFL00010) at 100 V for 2 h, cooled on ice.
- Air dry the blot.
- Wet the Immobilon®-FL PVDF membrane again in methanol for 30 sec.
- Wash membrane once with Milli-Q® water for 1 min.
- Block membrane with TBST (TBS + 0.1% Tween-20; TBS = 50 mM Tris, 150 mM NaCl, pH 7.4) containing 5% non-fat dry milk for 20 min at room temperature.
- Wash membrane 3× for 10 min with TBST.
- Incubate membrane with primary rabbit anti-iRhom2 antibody (antibodies-online, ABIN2846535, lot SA180724YE) diluted 1:1,000 in TBST containing 1% BSA overnight at 4°C.
- Wash membrane 3× for 10 min with TBST.
- Incubate membrane with secondary donkey anti-rabbit IgG (H+L), highly cross-adsorbed, Alexa Fluor™ Plus 800 (Thermo Fisher, A32808) diluted 1:100,000 in TBST containing 1% BSA for 1 h at room temperature.
- Wash membrane 1× for 10 min with TBST and 2× for 10 min with TBS.
- Detect stained proteins via fluorescence using the Odyssey 9120 imager system (LI-COR).
- Experimental Notes
- Human or murine iRhom2 (lanes 3 and 4) were overexpressed using the pMOWS vector via stable transduction with retroviruses produced by Phoenix Ampho cells, as described in Düsterhöft et al. 2021.
- iRhom1/iRhom2 double-knockout HEK293 cells were generated using the CRISPR-Cas9 system.
- No endogenous human iRhom2 was detected in the lysate of HEK293 cells (lane 1).
- Human iRhom2 was successfully detected in iRhom1/iRhom2 double-knockout HEK293 cells overexpressing human iRhom2 (lane 3).
- Overexpressed murine iRhom2 in iRhom1/iRhom2 double-knockout HEK293 cells was detected at substantially lower levels than overexpressed human iRhom2 (lane 4).
- Non-specific bands were also detected.
生效 #104634 (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 using ABIN2846535 to reveal iRhom2 (full length 110 kDa, cleaved 55 kDa) in lysates from HEK293 cells (1), HEK293 iRhom1/iRhom2 double knockout (dKO) (2), HEK293 iRhom1/iRhom2 dKO + overexpression of human iRhom2 wt (3), HEK293 iRhom1/iRhom2 dKO + overexpression of murine iRhom2 wt (4)]()
Western blot using ABIN2846535 to reveal iRhom2 (full length 110 kDa, cleaved 55 kDa) in lysates from HEK293 cells (1), HEK293 iRhom1/iRhom2 double knockout (dKO) (2), HEK293 iRhom1/iRhom2 dKO + overexpression of human iRhom2 wt (3), HEK293 iRhom1/iRhom2 dKO + overexpression of murine iRhom2 wt (4)
Full Methods -
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状态
- Liquid
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缓冲液
- Purified polyclonal antibody supplied in PBS with 0.09 % (W/V) sodium azide.
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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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储存条件
- 4 °C,-20 °C
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储存方法
- RHBDF2 Antibody (N-term) can be refrigerated at 2-8 °C for up to 6 months. For long term storage, place the at -20 °C.
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有效期
- 6 months
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- RHBDF2 (Rhomboid 5 Homolog 2 (RHBDF2))
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别名
- RHBDF2
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背景
- Not expected to have protease activity (By similarity).
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分子量
- 96686
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基因ID
- 79651
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NCBI登录号
- NP_001005498, NP_078875
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UniProt
- Q6PJF5
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途径
- Growth Factor Binding
抗原
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