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TIAM1 抗体 (T-Cell Lymphoma Invasion and Metastasis 1) (AA 1-378)

Details for Product anti-TIAM1 Antibody No. ABIN1982879, 供应商: Log in to see
抗原
  • TIAM1
  • tiam1
  • AI847750
  • D16Ium10
  • D16Ium10e
  • T-cell lymphoma invasion and metastasis 1
  • T-cell lymphoma invasion and metastasis 1a
  • T-cell lymphoma invasion and metastasis
  • T cell lymphoma invasion and metastasis 1
  • TIAM1
  • tiam1a
  • tiam1
  • LOC100338333
  • Tiam1
抗原表位
AA 1-378
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1
适用
小鼠
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29
19
2
2
2
2
1
1
宿主
绵羊
49
6
2
克隆类型
多克隆
标记
This TIAM1 antibody is un-conjugated
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3
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2
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2
1
1
1
1
1
1
1
1
1
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1
应用范围
Western Blotting (WB)
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9
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2
1
Supplier
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Available images

'独立验证'标志
抗原 TIAM1
Lot Number 2016090702
Method validated Western Blotting
Positive Control primary mouse microglia, MCF-7
Notes Passed. ABIN1982879 specifically recognizes TIAM1 in lysates from human MCF-7 cells.
Primary Antibody ABIN1982879
Secondary Antibody anti-goat/sheep HRP-conjugated antibodies (DakoCytomation, P0449, lot 00062104)
实验流程
  • Primary mouse microglia and human MCF-7 cells 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 plate or in RPMI 1640 (Invitrogen), supplemented with 6% inactivated foetal bovine serum (Gibco), GlutaMax (Gibco), penicillin and streptomycin (Gibco), at 37°C and 5% CO2 in a 10cm dish, respectively.
  • Lyse microglia in 25µl per well and MCF-7 cells in 500µl per dish cold lysis buffer (10mM Tris-HCl pH 8.0, 150mM NaCl, 1% Nonidet-P40, 1mM EDTA, 10mM NaF, 1mM Na3VO4, containing protease inhibitors (Sigma)).
  • Determine total protein content of the lysates using Bradford assay (Bio-Rad, 500-0006, lot 111832).
  • Denature 30µg total protein for 5min at 95°C in in 20µl Laemmli SDS sample buffer and subsequently separate them on a denaturing, freshly cast 7.5% SDS-PAGE for 1h at 140V.
  • Transfer proteins onto PVDF membrane (Pall Life Sciences, 75696G, lot TO3225) with a wet Western blotting system for 80min at 100V.
  • Block the membrane with TBST (TBS, 0.1% Tween) containing 5% milk ON at 4°C.
  • Incubation with primary sheep anti-TIAM1 antibody (antibodies-online, ABIN1982879, lot 2016090702) diluted 1:500 in TBST containing 5% milk for 1h at RT.
  • Wash membrane 5x 5min with TBST.
  • Incubation with anti-goat/sheep horseradish peroxidase-conjugated secondary antibodies (DakoCytomation, P0449, lot 00062104) 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 self-made ECL Western Blotting Detection Reagent with Curix RP2 Plus medical X-ray films (Agfa, ENKMV, lot 79040074) on an Agfa- Curix60-developer.
Experimental Notes The TIAM1 antibody ABIN1982879 reveals a protein of the expected molecular weight of 177kDa in human MCF-7 lysates as verified with a second anti-TIAM1 antibody. However, note that ABIN1982879 did not recognize Tiam1 in primary mouse microglia extracts.
Validation Images
Western Blotting validation image for anti-T-Cell Lymphoma Invasion and Metastasis 1 (TIAM1) (AA 1-378) antibody (ABIN1982879) ABIN1982879 was used to reveal TIAM1 in lysates from human MCF-7 (lane 2) cells and m...
免疫原 AA 1-378
亚型 IgG
产品特性 Polyclonal Sheep anti Mouse TIAM1
别名 TIAM1 (TIAM1 Antibody 摘要)
UniProt Q13009
研究领域 Signaling
应用备注 Optimal dilutions should be determined by each laboratory for each application.
限制 仅限研究用
状态 Lyophilized
溶解方式 100 µL of PBS, pH 7.4
浓度 500 µg/mL
缓冲液 Lyophilized from a solution containing PBS, pH 7.4
储存条件 -20 °C