ALPK1 抗体 (N-Term)
(1 validation)-
- 抗原 See all ALPK1 抗体
- ALPK1 (alpha-Kinase 1 (ALPK1))
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抗原表位
- N-Term
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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 ALPK1 antibody is un-conjugated
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应用范围
- Western Blotting (WB), ELISA, Immunohistochemistry (IHC), Immunofluorescence (IF)
- 特异性
- α-protein Kinase 1 Polyclonal Antibody detects endogenous levels of α-protein Kinase 1 protein.
- 产品特性
- Rabbit Polyclonal to α-protein Kinase 1.
- 纯化方法
- The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen.
- 免疫原
- Synthesized peptide derived from the N-terminal region of human alpha-protein Kinase 1.
- 亚型
- IgG
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anti-alpha-Kinase 1 (ALPK1) (AA 1-1085) antibody
ALPK1 适用: 人 WB 宿主: 小鼠 Polyclonal unconjugated
anti-alpha-Kinase 1 (ALPK1) (C-Term) antibodyALPK1 适用: 人 ELISA, IHC 宿主: 兔 Polyclonal unconjugated
anti-alpha-Kinase 1 (ALPK1) (Internal Region) antibodyALPK1 适用: 人, 小鼠 WB, ELISA, IHC, IF, ICC 宿主: 兔 Polyclonal unconjugated
anti-alpha-Kinase 1 (ALPK1) (N-Term) antibodyALPK1 适用: 人 WB, IHC, IF, IC 宿主: 兔 Polyclonal unconjugated
anti-alpha-Kinase 1 (ALPK1) (AA 11-60) antibodyALPK1 适用: 人, 小鼠 WB, ELISA, IHC, IHC (p) 宿主: 兔 Polyclonal unconjugated
anti-alpha-Kinase 1 (ALPK1) (AA 11-60) antibodyALPK1 适用: 人, 小鼠 WB, ELISA 宿主: 兔 Polyclonal unconjugated
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- 应用备注
- WB 1:500-1:2000, IHC 1:100-1:300, IF 1:200-1:1000, ELISA 1:20000,
- 限制
- 仅限研究用
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- by
- Institute of Experimental Internal Medicine, Otto von Guericke Universität Magdeburg
- No.
- #103932
- 日期
- 2019.08.26
- 抗原
- ALPK1
- Lot Number
- 63660
- Method validated
- Western Blotting
- Positive Control
Recombinant c-Myc-tagged hALPK1 overexpression in AGS KOhALPK1
- Negative Control
AGS KOhALPK1
- Notes
ABIN3187581 detects overexpressed hALPK1 but not the endogenous protein in AGS cells.
- Primary Antibody
- ABIN3187581
- Secondary Antibody
- donkey anti-rabbit HRP antibody (Jackson ImmunResearch, 711-036-152, lot 140655)
- Full Protocol
- Grow AGS (ATCC, CRL-1739 ) in RPMI 1640 (Gibco) supplemented with 10% FCS (Gibco) and no antibiotics, at 37°C and 5% CO2 in 4ml on a 60mm dish to 70% confluency.
- Transfect cells with 5µg phALPK1 (Origene) using Metafectene Pro (Biontex) following the manufacturer´s instructions.
- Lyse 1.6x106 cells in 70µl per dish of ice-cold M-PER (Pierce) supplemented with Halt Protease and Phosphatase Inhibitor Cocktail (ThermoFisher, 78440) after two PBS washes.
- Determine total protein content of the lysates using BCA (ThermoFisher).
- Denature 200µg of total protein for 5min at 95°C in 100µl and subsequently separate 20µg or 40µg on a freshly cast denaturing 10% polyacrylamide gel in a Bio-Rad Protean 3 for 15min at 100V and 1.5h at 140V.
- Transfer proteins onto membrane 0.45µm PVDF (Millipore, IPVH00010) with a Bio-Rad Western blotting system for 2h at 100V constant.
- Block the membrane with TBS-T and 5% skim milk (Roth) for 1 at RT.
- Incubation with primary rabbit-anti ALPK1 antibody (antibodies-online, ABIN3187581, lot 63660) diluted 1:1000 in TBS-T containing BSA (AppliChem, A1391, 0500) ON at 4°C.
- Wash membrane 3x for 5, 10, 10 minutes with TBS-T.
- Incubation with secondary donkey anti-rabbit HRP antibody (Jackson ImmunResearch, 711-036-152, lot 140655) diluted 1:10000 in TBS-T and 5% skim milk (Roth) for 1h at RT.
- Wash membrane 3x for 10min with TBS-T.
- Reveal protein bands using Immobilon Western (Millipore, WBKLS0500, lot 1827003) using ChemoCAM (INTAS) as gel documentation system.
- Experimental Notes
ABIN3187581 detected very strong two unspecific bands running at approximately 60kDa and 95kDa. This was visible incubating the antibody in milk or albumin. In neither milk nor albumin the antibody was able to detect the endogenous hALPK1 present in AGS cells. Incubation in albumin but not milk lead to the detection of recombinant overexpressed hALPK1 as confirmed by c-Myc detection (SCBT, sc-40) of the recombinant protein. Another antibody available in our laboratory (Proteintech Europe 19107-1-AP, lot 00010221) also detects the overexpressed hALPK1 (~135kDa) but not the endogenous hALPK1.
It is possible that the amount of ALPK1 in our cell preparation is low. Other cell lysate might be better suited. From my data, I can conclude that this antibody detects overexpressed hALPK1.
Because no antibody detecting the endogenous hALPK1 is available to me, I cannot confirm the knockout in our cell lines, but on the functional level, we have indications for a successful knockout.
生效 #103932 (Western Blotting)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![]()
Full Methods -
- 状态
- Liquid
- 浓度
- 1 mg/mL
- 缓冲液
- Liquid in PBS containing 50 % glycerol, 0.5 % BSA and 0.02 % sodium azide.
- 储存液
- Sodium azide
- 注意事项
- This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- 注意事项
- Avoid repeated freeze/thaw cycles.
- 储存条件
- -20 °C
- 储存方法
- Store at -20°C.
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