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Figure 1. Western blot analysis of MAP3K8 using anti-MAP3K8 antibody (PB10007).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human HEPG2 whole cell lysates,
Lane 2: human HELA whole cell lysates,
Lane 3: human A431 whole cell lysates,
Lane 4: human A549 whole cell lysates,
Lane 5: human THP-1 whole cell lysates,
Lane 6: human Daudi whole cell lysates,
Lane 7: rat liver tissue lysates,
Lane 8: mouse liver tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MAP3K8 antigen affinity purified polyclonal antibody (Catalog # PB10007) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MAP3K8 at approximately 53KD. The expected band size for MAP3K8 is at 53KD.
Figure 2. IF analysis of MAP3K8 using anti-MAP3K8 antibody (PB10007).
MAP3K8 was detected in immunocytochemical section of HepG2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-MAP3K8 Antibody (PB10007) overnight at 4°C. DyLightR488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Figure 3. Flow Cytometry analysis of U937 cells using anti-MAP3K8 antibody (PB10007).
Overlay histogram showing U937 cells stained with PB10007 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-MAP3K8 Antibody (PB10007, 1μg/1x106 cells) for 30 min at 20°C. DyLightR488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Figure 4. IHC analysis of MAP3K8 using anti-MAP3K8 antibody (PB10007).
MAP3K8 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MAP3K8 Antibody (PB10007) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Figure 5. IHC analysis of MAP3K8 using anti-MAP3K8 antibody (PB10007).
MAP3K8 was detected in paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MAP3K8 Antibody (PB10007) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Figure 6. IHC analysis of MAP3K8 using anti-MAP3K8 antibody (PB10007).
MAP3K8 was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MAP3K8 Antibody (PB10007) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Figure 7. IHC analysis of MAP3K8 using anti-MAP3K8 antibody (PB10007).
MAP3K8 was detected in paraffin-embedded section of human pancreatic cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MAP3K8 Antibody (PB10007) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Figure 8. IHC analysis of MAP3K8 using anti-MAP3K8 antibody (PB10007).
MAP3K8 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-MAP3K8 Antibody (PB10007) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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Figure 1. Western blot analysis of MAP3K8 using anti-MAP3K8 antibody (PB10007).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human HEPG2 whole cell lysates,
Lane 2: human HELA whole cell lysates,
Lane 3: human A431 whole cell lysates,
Lane 4: human A549 whole cell lysates,
Lane 5: human THP-1 whole cell lysates,
Lane 6: human Daudi whole cell lysates,
Lane 7: rat liver tissue lysates,
Lane 8: mouse liver tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-MAP3K8 antigen affinity purified polyclonal antibody (Catalog # PB10007) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for MAP3K8 at approximately 53KD. The expected band size for MAP3K8 is at 53KD.
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| 別品名 |
Mitogen-activated protein kinase kinase kinase 8;2.7.11.25;Cancer Osaka thyroid oncogene;Proto-oncogene c-Cot;Serine/threonine-protein kinase cot;Tumor progression locus 2;TPL-2;MAP3K8;COT, ESTF;
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| 種由来 |
Human
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| 交差種 |
Human
Rat
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| 適用 |
Western Blot
Immunohistochemistry
Immuno Fluorescence
Immunocytochemistry (cell)
Flow Cytometry
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| 免疫動物 |
Rabbit
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| 抗体クラス |
IgG
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| 標識物 |
Phycoerythrin
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| 精製度 |
Affinity Purified
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| GENE ID |
1326
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| Accession No.(Gene/Protein) |
P41279
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| Gene Symbol |
MAP3K8
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| 分子量 |
52925 MW
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| 概要 |
Boster Bio Anti-MAP3K8 Antibody Picoband® catalog # PB10007. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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| 参考文献 |
1. "Entrez Gene: MAP3K8 mitogen-activated protein kinase kinase kinase 8".2. Chan AM, Chedid M, McGovern ES, Popescu NC, Miki T, Aaronson SA (May 1993). "Expression cDNA cloning of a serine kinase transforming gene". Oncogene 8 (5): 1329-33.3. Miyoshi J, Higashi T, Mukai H, Ohuchi T, Kakunaga T (Aug 1991). "Structure and transforming potential of the human cot oncogene encoding a putative protein kinase". Molecular and Cellular Biology 11 (8): 4088-96.
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| メーカー |
品番 |
包装 |
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BBT
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PB10007-PE
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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 |
1週間程度
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| 保存温度 |
-２０℃
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