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Product Info Summary
| SKU: | A00185 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-TRAF6 Antibody Picoband®
SKU/Catalog Number
A00185
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-TRAF6 Antibody Picoband® catalog # A00185. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with 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
Store 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 freeze-thaw cycles.
Cite This Product
Anti-TRAF6 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00185)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived mouse TRAF6 recombinant protein (Position: Q14-D512).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00185 is reactive to Traf6 in Mouse, Rat
Observed Molecular Weight
60-65 kDa
Calculated molecular weight
60.1 kDa
Background of Traf6
TNF receptor-associated factor 6, also called E3 ubiquitin-protein ligase TRAF6 or RNF85, is a TRAF human protein. The protein encoded by this gene is a member of the TNF receptor associated factor (TRAF) protein family. TRAF proteins are associated with, and mediate signal transduction from, members of the TNF receptor superfamily. This gene is mapped to 11p12. This protein mediates signaling from members of the TNF receptor superfamily as well as the Toll/IL-1 family. Signals from receptors such as CD40, TNFSF11/RANCE and IL-1 have been shown to be mediated by this protein. This protein also interacts with various protein kinases including IRAK1/IRAK, SRC and PKCzeta, which provides a link between distinct signaling pathways. This protein functions as a signal transducer in the NF-kappaB pathway that activates IkappaB kinase (IKK) in response to proinflammatory cytokines. The interaction of this protein with UBE2N/UBC13, and UBE2V1/UEV1A, which are ubiquitin conjugating enzymes catalyzing the formation of polyubiquitin chains, has been found to be required for IKK activation by this protein.
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
A00185 is guaranteed for ELISA, Flow Cytometry, IHC, 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, 0.25-0.5μg/ml, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Mouse, Rat
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Mouse
ELISA, 0.1-0.5μg/ml, -
"
Positive Control
WB: rat RH35 whole cell, mouse thymus tissue, mouse spleen tissue, mouse RAW2467 whole cell
IHC: rat intestine tissue, mouse intestine tissue
FCM: HEPA 1-6 cell
Validation Images & Assay Conditions
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Western blot analysis of TRAF6 using anti-TRAF6 antibody (A00185).
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: rat RH35 whole cell lysates,
Lane 2: mouse thymus tissue lysates,
Lane 3: mouse spleen tissue lysates,
Lane 4: mouse RAW246.7 whole cell 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-TRAF6 antigen affinity purified polyclonal antibody (Catalog # A00185) 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 1:10000 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 TRAF6 at approximately 60-65KD. The expected band size for TRAF6 is at 60KD.
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IHC analysis of TRAF6 using anti-TRAF6 antibody (A00185).
TRAF6 was detected in paraffin-embedded section of rat intestine 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-TRAF6 Antibody (A00185) 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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IHC analysis of TRAF6 using anti-TRAF6 antibody (A00185).
TRAF6 was detected in paraffin-embedded section of mouse intestine 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-TRAF6 Antibody (A00185) 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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Flow Cytometry analysis of HEPA 1-6 cells using anti-TRAF6 antibody (A00185).
Overlay histogram showing HEPA 1-6 cells stained with A00185 (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-TRAF6 Antibody (A00185, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 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.
Specific Publications For Anti-TRAF6 Antibody Picoband® (A00185)
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1 Customer Q&As for Anti-TRAF6 Antibody Picoband®
Question
We are currently using anti-TRAF6 antibody A00185 for mouse tissue, and we are happy with the Flow Cytometry results. The species of reactivity given in the datasheet says mouse, rat. Is it likely that the antibody can work on horse tissues as well?
Verified Customer
Verified customer
Asked: 2019-06-26
Answer
The anti-TRAF6 antibody (A00185) has not been tested for cross reactivity specifically with horse tissues, but there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in horse you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-06-26
