RHOT1 抗体 (N-Term)
(1 validation)Quick Overview for RHOT1 抗体 (N-Term) (ABIN635090)
抗原
See all RHOT1 抗体适用
宿主
克隆类型
标记
应用范围
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抗原表位
- N-Term
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特异性
- RHOT1 antibody was raised against the N terminal of RHOT1
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纯化方法
- Affinity purified
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免疫原
- RHOT1 antibody was raised using the N terminal of RHOT1 corresponding to a region with amino acids EMKPACIKALTRIFKISDQDNDGTLNDAELNFFQRICFNTPLAPQALEDV
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anti-Ras Homolog Gene Family, Member T1 (RHOT1) (N-Term) antibody
RHOT1 适用: 人, 小鼠, 大鼠, 犬, Cow, 豚鼠, 马, 兔, 斑马鱼 WB, IHC 宿主: 兔 Polyclonal unconjugated
anti-Ras Homolog Gene Family, Member T1 (RHOT1) (AA 483-580) antibodyRHOT1 适用: 人 WB, ELISA 宿主: 小鼠 Monoclonal 4H4 unconjugated
anti-Ras Homolog Gene Family, Member T1 (RHOT1) (N-Term) antibodyRHOT1 适用: 人, 小鼠, 大鼠, 犬, Cow, 豚鼠, 马, 兔, 斑马鱼 WB 宿主: 兔 Polyclonal unconjugated
anti-Ras Homolog Gene Family, Member T1 (RHOT1) (N-Term) antibodyRHOT1 适用: 人, 小鼠, 大鼠, Cow, 豚鼠, 马, 兔, Bat, 小鸡, 猴, Pig WB, IHC, IHC (p) 宿主: 兔 Polyclonal unconjugated
anti-Ras Homolog Gene Family, Member T1 (RHOT1) (N-Term) antibodyRHOT1 适用: 人, 小鼠, 犬 WB, IHC 宿主: 兔 Polyclonal unconjugated
anti-Ras Homolog Gene Family, Member T1 (RHOT1) (AA 483-580) antibodyRHOT1 适用: 人 WB, ELISA, IF 宿主: 小鼠 Monoclonal 1A12 unconjugated
anti-Ras Homolog Gene Family, Member T1 (RHOT1) (AA 483-580) antibodyRHOT1 适用: 人 WB, ELISA 宿主: 小鼠 Polyclonal unconjugated
anti-Ras Homolog Gene Family, Member T1 (RHOT1) (AA 1-659) antibodyRHOT1 适用: 人 WB 宿主: 小鼠 Polyclonal unconjugated
anti-Ras Homolog Gene Family, Member T1 (RHOT1) (AA 396-445) antibodyRHOT1 适用: 人, 小鼠, 大鼠, 犬, Cow, 豚鼠, 马, 兔, Bat, 小鸡, 猴, Pig WB 宿主: 兔 Polyclonal unconjugated
anti-Ras Homolog Gene Family, Member T1 (RHOT1) (AA 180-229) antibodyRHOT1 适用: 人, 小鼠, 大鼠, 犬, Cow, 豚鼠, 马, 兔, Bat, 小鸡, 猴, Pig WB 宿主: 兔 Polyclonal unconjugated
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应用备注
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WB: 1 µg/mL
Optimal conditions should be determined by the investigator. -
说明
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RHOT1 Blocking Peptide, (ABIN937971), is also available for use as a blocking control in assays to test for specificity of this RHOT1 antibody
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限制
- 仅限研究用
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- by
- Department of Biochemistry, University of Utah, Shaw lab
- No.
- #100007
- 日期
- 2015.09.12
- 抗原
- Ras Homolog Gene Family, Member T1 (RHOT1) (N-Term)
- Lot Number
- Method validated
- Western Blotting
- Positive Control
- 100ng Recombinant Miro1; 30ug Mitochondria isolated from Miro1 WT MEFs and Miro1 KO MEFs expressing Myc-Miro1; in house purification and isolation Isolated mitochondria from immortalized Miro1 WT MEFs rMiro1, mtMiro1 wt MEFs, mtMiro1 KO MEFs plus Venus-Miro1
- Negative Control
- 100ng Recombinant Miro2, 30ug Mitochondria isolated from Miro1 KO MEFs and Miro1 KO MEFs expressing Myc-Miro2; in house purification and isolation
- Notes
- ABIN635090 is specific for Miro1 without cross-reaction with Miro2.
- Primary Antibody
- ABIN635090
- Secondary Antibody
- Goat-anti-rabbit IgG HRP conjugated (Sigma A0545-1M2 - 057K4802) 1:2000 in TBS + 0.05% Tween-20 + 5% non-fat dry milk
- Full Protocol
- Transfection of immortalized Miro1-/- MEFs with Myc-Miro1-V1 or Myc-Miro2-V1 (Published in Nguyen et al., 2014 (PNAS, 10.1073/pnas.1402449111))
- Isolation of mitochondria from immortalized Miro1+/+, Miro1-/-, Miro1-/- + Myc-Miro1 and Miro1-/- + Myc-Miro2 MEFs
- Determine protein concentration and snap freeze mitochondria in liquid nitrogen
Day 1
- Thaw 30ug mitochondria per lane on ice
- Thaw 100ng recombinant Miro1 and Miro2 per lane on ice (in house purification)
- Add 2x Laemmli buffer + 2% BME to samples and denature for 5 min at 95C
- Load 30ug/lane mitochondria and 100ng/lane recombinant protein onto 12.5 % SDS-PAGE
- Fermentas PageRuler Plus Prestained (2ul) - Lane 7 / 14 / 21
- Recombinant Miro1- Lane 1 / 8 / 15
- Recombinant Miro2- Lane 2 / 9 / 16
- Miro1+/+ mitochondria- Lane 3 / 10 / 17
- Miro1-/- mitochondria- Lane 4 / 11 / 18 / 22
- Miro1-/- + Myc-Miro1-V1 mitochondria- Lane 5 / 12 / 19 / 23
- Miro1-/- + Myc-Miro2-V1 mitochondria- Lane 6 / 13 / 20 / 24
- Run gel 40mA 180V for 3h
- Semi-dry transfer to PVDF 250mA 15V for 2h
- Block with TBS + 3% BSA for 1h at RT
Lanes Antibody Host Dilution In 1-6 Anti-Miro1 Other company - - - 8-13 Anti-Miro1 ABIN634527v rabbit 1:1000 TBST + 3% BSA 15-20 Anti-Miro1 ABIN635090 rabbit 1:1000 TBST + 3% BSA 22-24 Anti-Myc - - - All lanes Anti-Cyt c - - - Anti-PRX3 - - - - Day 2
- 3x washing with TBS + 0.05% Tween-20 for 5'
- 2nd AB for 1h at RT
- Goat-anti-rabbit IgG HRP (Sigma A0545-1M2 - 057K4802)
- 1:2000 in TBS + 0.05% Tween-20 + 5% non-fat dry milk3x washing with TBS for 10'
- Detection with homemade ECL prepared according to Haan and Behrmann, 2007 (J Immunol Methods, 10.1016/j.jim.2006.07.027) on a Bio-Rad imaging system
- Experimental Notes
生效 #100007 (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 customer validation ABIN635090. Refer to the protocol section for more information.]()
Western blot customer validation ABIN635090. Refer to the protocol section for more information.
Full Methods -
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状态
- Lyophilized
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溶解方式
- Lyophilized powder. Add distilled water for a 1 mg/mL concentration of RHOT1 antibody in PBS
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浓度
- Lot specific
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缓冲液
- PBS
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注意事项
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Avoid repeated freeze/thaw cycles.
Dilute only prior to immediate use. -
储存条件
- 4 °C/-20 °C
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储存方法
- Store at 2-8 °C for short periods. For longer periods of storage, store at -20 °C.
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- RHOT1 (Ras Homolog Gene Family, Member T1 (RHOT1))
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别名
- RHOT1
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背景
- RHOT1 is mitochondrial GTPase involved in mitochondrial trafficking. It is probably involved in control of anterograde transport of mitochondria and their subcellular distribution.
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分子量
- 75 kDa (MW of target protein)
抗原
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