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Product Info Summary
| SKU: | A00929-3 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-ASK1/MAP3K5 Antibody Picoband®
SKU/Catalog Number
A00929-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ASK1/MAP3K5 Antibody Picoband® catalog # A00929-3. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, 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.
Storage & Handling
At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.
Cite This Product
Anti-ASK1/MAP3K5 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00929-3)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human ASK1/MAP3K5 recombinant protein (Position: A178-K1112).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00929-3 is reactive to MAP3K5 in Human, Mouse, Rat
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
Observed Molecular Weight
155 kDa
Calculated molecular weight
27904 MW
Background of ASK1
Apoptosis signal-regulating kinase 1 (ASK1) also known as mitogen-activated protein kinase 5 (MAP3K5) is a member of MAP kinase family and as such a part of mitogen-activated protein kinase pathway. Mitogen-activated protein kinase (MAPK) signaling cascades include MAPK or extracellular signal-regulated kinase (ERK), MAPK kinase (MKK or MEK), and MAPK kinase kinase (MAPKKK or MEKK). MAPKK kinase/MEKK phosphorylates and activates its downstream protein kinase, MAPK kinase/MEK, which in turn activates MAPK. The kinases of these signaling cascades are highly conserved, and homologs exist in yeast, Drosophila, and mammalian cells. MAPKKK5 contains 1,374 amino acids with all 11 kinase subdomains. Northern blot analysis shows that MAPKKK5 transcript is abundantly expressed in human heart and pancreas. The MAPKKK5 protein phosphorylates and activates MKK4 (aliases SERK1, MAPKK4) in vitro, and activates c-Jun N-terminal kinase (JNK)/stress-activated protein kinase (SAPK) during transient expression in COS and 293 cells; MAPKKK5 does not activate MAPK/ERK.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00929-3 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human RT4 whole cell, human HEL whole cell, human HL-60 whole cell, human THP-1 whole cell, rat liver tissue, mouse liver tissue,
IHC: human lung cancer tissue, human spleen tissue
ICC/IF: MCF-7 cell
FCM: Raji cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of ASK1/MAP3K5 using anti-ASK1/MAP3K5 antibody (A00929-3).
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 30 ug of sample under reducing conditions.
Lane 1: human RT4 whole cell lysates,
Lane 2: human HEL whole cell lysates,
Lane 3: human HL-60 whole cell lysates,
Lane 4: human THP-1 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: mouse liver tissue lysates,
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-ASK1/MAP3K5 antigen affinity purified polyclonal antibody (Catalog # A00929-3) 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 ASK1/MAP3K5 at approximately 155 kDa. The expected band size for ASK1/MAP3K5 is at 155 kDa.
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Figure 2. IHC analysis of ASK1/MAP3K5 using anti-ASK1/MAP3K5 antibody (A00929-3).
ASK1/MAP3K5 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-ASK1/MAP3K5 Antibody (A00929-3) 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 3. IHC analysis of ASK1/MAP3K5 using anti-ASK1/MAP3K5 antibody (A00929-3).
ASK1/MAP3K5 was detected in a paraffin-embedded section of human spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-ASK1/MAP3K5 Antibody (A00929-3) 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 4. IF analysis of ASK1/MAP3K5 using anti-ASK1/MAP3K5 antibody (A00929-3).
ASK1/MAP3K5 was detected in an immunocytochemical section of MCF-7 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 5 μg/mL rabbit anti-ASK1/MAP3K5 Antibody (A00929-3) overnight at 4°C. DyLight®488 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.
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Figure 5. Flow Cytometry analysis of Raji cells using anti-ASK1/MAP3K5 antibody (A00929-3).
Overlay histogram showing Raji cells stained with A00929-3 (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-ASK1/MAP3K5 Antibody (A00929-3, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For MAP3K5 (Source: Uniprot.org, NCBI)
Gene Name
MAP3K5
Full Name
Mitogen-activated protein kinase kinase kinase 5
Weight
27904 MW
Superfamily
protein kinase superfamily
Alternative Names
Insulin-like growth factor-binding protein 1; IBP-1; IGF-binding protein 1; IGFBP-1; Igfbp1; Igfbp-1; MAP3K5 ASK1, MAPKKK5, MEKK5 mitogen-activated protein kinase kinase kinase 5 mitogen-activated protein kinase kinase kinase 5|ASK-1|MAP/ERK kinase kinase 5|MAPK/ERK kinase kinase 5|MEK kinase 5|MEKK 5|apoptosis signal-regulating kinase 1
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on MAP3K5, check out the MAP3K5 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for MAP3K5: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-ASK1/MAP3K5 Antibody Picoband® (A00929-3)
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