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Product Info Summary
| SKU: | A00284-4 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, WB |
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Product info
Product Name
Anti-NF-κB p65/RELA Antibody Picoband®
SKU/Catalog Number
A00284-4
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-NF-κB p65/RELA Antibody Picoband® catalog # A00284-4. Tested in ELISA, WB, Flow Cytometry 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-NF-κB p65/RELA Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00284-4)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
IgG
Immunogen
E.coli-derived human NF-κB p65/RELA recombinant protein (Position: Q132-S551). Human RELA shares 85.8% amino acid (aa) sequence identity with mouse RELA.
Cross-reactivity
No cross reactivity with other proteins.
Reactive Species
A00284-4 is reactive to RELA in Human, Mouse, Rat
Observed Molecular Weight
65 kDa
Calculated molecular weight
60.2 kDa
Background of RELA
Transcription factor p65, also known as NFKB3 or NF-kB p65, is a protein that in humans is encoded by the RELA gene. It is mapped to 11q13.1. NFKB is an essential transcription factor complex involved in all types of cellular processes, including cellular metabolism, chemotaxis, etc, and it may play a role in inflammatory conditions of the peripheral nervous system. Phosphorylation and acetylation of NFKB3 are crucial post-translational modifications required for NFKB activation. It has also been shown to modulate immune responses, and activation of NFKB3 is positively associated with multiple types of cancer. In addition to that, NFKB3 antagonizes TNFR1-JNK proliferative signals in epidermis and plays a nonredundant role in restraining epidermal growth.
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
A00284-4 is guaranteed for ELISA, Flow Cytometry, 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, 0.1-0.25 μg/ml, Human, Mouse, Rat
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human Hela whole cell, human HepG2 whole cell, human Caco-2 whole cell, human Raji whole cell, rat PC-12 whole cell, rat C6 whole cell tissue, mouse NIH/3T3 whole cell, mouse Ana-1 whole cell
FCM: Daudi cell
Validation Images & Assay Conditions
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Western blot analysis of NF-ΚB P6/RELA using anti-NF-ΚB P6/RELA antibody (A00284-4).
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 Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human Caco-2 whole cell lysates,
Lane 4: human Raji whole cell lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: rat C6 whole cell tissue lysates,
Lane 7: mouse NIH/3T3 whole cell lysates,
Lane 8: mouse Ana-1 whole cell 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-NF-ΚB P6/RELA antigen affinity purified polyclonal antibody (Catalog # A00284-4) at 0.25 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 NF-ΚB P6/RELA at approximately 65 kDa. The expected band size for NF-ΚB P6/RELA is at 60 kDa.
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Flow Cytometry analysis of Daudi cells using anti-NF-ΚB P6/RELA antibody (A00284-4).
Overlay histogram showing Daudi cells stained with A00284-4 (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-NF-ΚB P6/RELA Antibody (A00284-4, 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 (Red line) was also used as a control.
Specific Publications For Anti-NF-κB p65/RELA Antibody Picoband® (A00284-4)
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