TIAM1 抗体 (AA 1-378)
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北京 101111
Quick Overview for TIAM1 抗体 (AA 1-378) (ABIN1982879)
抗原
See all TIAM1 抗体适用
宿主
克隆类型
标记
应用范围
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抗原表位
- AA 1-378
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产品特性
- Polyclonal Sheep anti Mouse TIAM1
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免疫原
- AA 1-378
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亚型
- IgG
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anti-T-Cell Lymphoma Invasion and Metastasis 1 (TIAM1) (AA 1392-1591) antibody
TIAM1 适用: 人 WB, IHC 宿主: 兔 Polyclonal unconjugated
anti-T-Cell Lymphoma Invasion and Metastasis 1 (TIAM1) (AA 52-224) antibodyTIAM1 适用: 人 ELISA, IHC, IF 宿主: 兔 Polyclonal unconjugated
anti-T-Cell Lymphoma Invasion and Metastasis 1 (TIAM1) (AA 1478-1506), (C-Term) antibodyTIAM1 适用: 人 WB 宿主: 兔 Polyclonal RB41848 unconjugated
anti-T-Cell Lymphoma Invasion and Metastasis 1 (TIAM1) (C-Term) antibodyVerified TIAM1 适用: 人 ELISA, IF, FACS 宿主: 山羊 Polyclonal unconjugated
anti-T-Cell Lymphoma Invasion and Metastasis 1 (TIAM1) (AA 1478-1506) antibodyTIAM1 适用: 人 WB, IHC, IHC (p) 宿主: 兔 Polyclonal unconjugated
anti-T-Cell Lymphoma Invasion and Metastasis 1 (TIAM1) (AA 1261-1397) antibodyTIAM1 适用: 人 WB, IHC, IP, ICC 宿主: 兔 Polyclonal unconjugated
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应用备注
- Optimal dilutions should be determined by each laboratory for each application.
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限制
- 仅限研究用
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- by
- Instituto de Parasitología y Biomedicina López-Neyra
- No.
- #100102
- 日期
- 2016.12.23
- 抗原
- TIAM1
- Lot Number
- 2016090702
- Method validated
- Western Blotting
- Positive Control
- primary mouse microglia, MCF-7
- Negative Control
- 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)
- Full Protocol
- 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.
生效 #100102 (Western Blotting)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![ABIN1982879 was used to reveal TIAM1 in lysates from human MCF-7 (lane 2) cells and mouse primary mouse microglia (lane 1) as described in the protocol.]()
ABIN1982879 was used to reveal TIAM1 in lysates from human MCF-7 (lane 2) cells and mouse primary mouse microglia (lane 1) as described in the protocol.
Full Methods -
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状态
- Lyophilized
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溶解方式
- 100 µL of PBS, pH 7.4
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浓度
- 500 µg/mL
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缓冲液
- Lyophilized from a solution containing PBS, pH 7.4
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储存条件
- -20 °C
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- TIAM1 (T-Cell Lymphoma Invasion and Metastasis 1 (TIAM1))
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别名
- TIAM1
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UniProt
- Q13009
抗原
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