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Rockland Rabbit anti-Transferrin (109-4134, lot 3033, green), Goat Anti-Alpha-1-Anti-Trypsin (100-101-147, lot 5842), and Mouse Anti-GST (200-301-200, lot 24882) were used in a multiplex system to detect target proteins under reducing (R) conditions (+4% BME) in albumin depleted human serum with 320ng of added GST. Sample was run by SDS-PAGE, transferred to 0.2 um PVDF using the BioRad Trans-Blot Turbo and blocked in 2.5% Blotto, 2.5% BSA, 0.02% Tween overnight at 4°C. Membrane was probed with three primary antibodies at 1:1000 dilution (in MB-070 overnight at 4°C). Detection shown was using DyLight549 Donkey anti-Rabbit IgG (611-742-127, lot 21100, shown as green), DyLight 488 Donkey anti-Mouse IgG (610-741-124, lot 21095, shown as blue), and DyLight 649 Donkey anti-Goat IgG (605-743-125, lot 20834, shown as red) at 1:10000 (in MB-070 30 min RT). Blots were washed, rinsed in methanol, dried and Images were collected using the BioRad VersaDoc System.
Western Blot of Anti-Rabbit IgG (H&L) (DONKEY) Antibody (Min X Bv Ch Gt GP Ham Hs Hu Ms Rt & Sh Serum Proteins) (p/n 611-701-127). Lane M: 3 μl Molecular Ladder. Lane 1: Rabbit IgG whole molecule (p/n 011-0102). Lane 2: Rabbit IgG F(ab) Fragment (p/n 011-0105). Lane 3: Rabbit IgG F(c) Fragment (p/n 010-0103). Lane 4: Rabbit IgM Whole Molecule (p/n 011-0107). Lane 5: Normal Rabbit Serum (p/n B309). All samples were reduced. Load: 50 ng of IgG, F(ab), F(c) and Serum, 25 ng of IgM. Block: MB-070 for 30 min at RT. Primary Antibody: Anti-Rabbit IgG (H&L) (DONKEY) Antibody (Min X Bv Ch Gt GP Ham Hs Hu Ms Rt & Sh Serum Proteins) (p/n 611-701-127) 1:7,500 for 60 min at RT. Secondary antibody: Anti-Donkey IgG (GOAT) Peroxidase Conjugated Antibody (p/n 616-1302) 1:40,000 in MB-070 for 30 min at RT. Predicted/Observed Size: 25 and 50 kDa for Rabbit IgG and Serum, 25 kDa for F(c) and F(ab), 70 and 23 kDa for IgM. Rabbit F(c) migrates slightly higher.
DyLightTM dyes can be used for two-color Western Blot detection with low background and high signal.？ Anti-tubulin was detected using a DyLightTM 549 conjugate.？ Anti-TNFa was detected using a DyLightTM 649 conjugate. The image was captured using the TyphoonTM 9410 Imaging System.
Properties of DyLightTM Conjugates.
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Rockland Rabbit anti-Transferrin (109-4134, lot 3033, green), Goat Anti-Alpha-1-Anti-Trypsin (100-101-147, lot 5842), and Mouse Anti-GST (200-301-200, lot 24882) were used in a multiplex system to detect target proteins under reducing (R) conditions (+4% BME) in albumin depleted human serum with 320ng of added GST. Sample was run by SDS-PAGE, transferred to 0.2 um PVDF using the BioRad Trans-Blot Turbo and blocked in 2.5% Blotto, 2.5% BSA, 0.02% Tween overnight at 4°C. Membrane was probed with three primary antibodies at 1:1000 dilution (in MB-070 overnight at 4°C). Detection shown was using DyLight549 Donkey anti-Rabbit IgG (611-742-127, lot 21100, shown as green), DyLight 488 Donkey anti-Mouse IgG (610-741-124, lot 21095, shown as blue), and DyLight 649 Donkey anti-Goat IgG (605-743-125, lot 20834, shown as red) at 1:10000 (in MB-070 30 min RT). Blots were washed, rinsed in methanol, dried and Images were collected using the BioRad VersaDoc System.
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| 別品名 |
Donkey Anti-Rabbit IgG Antibody DyLight 549TM Conjugated, Donkey Anti Rabbit IgG DyLight 549TM Conjugated Antibody
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| 交差種 |
Rabbit
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| 非交差(吸収処理)種 |
Human
Mouse
Rat
Bovine
Chicken
Sheep
Goat
Guinea Pig
Hamster
Equine
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| 適用 |
Western Blot
Dot Blot
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| 免疫動物 |
Donkey
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| 標識物 |
DyLightTM 549
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| 精製度 |
Affinity Purified
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| 参考文献 |
[[Cosmo Reference Collection]][Pub Med ID]27504635,23958595,23260141,29767252,28893621,31626844,34520793
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| [注意事項] |
濃度はロットによって異なる可能性があります。メーカーDS及びCoAからご確認ください。
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| メーカー |
品番 |
包装 |
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RKL
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611-742-127
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100 UG
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※表示価格について
| 当社在庫 |
なし
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| 納期目安 |
約10日
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| 法規制 |
毒
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| 保存温度 |
４℃
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