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Product Info Summary
| SKU: | A00284-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-NF-kB p65/RELA Antibody Picoband®
SKU/Catalog Number
A00284-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-NF-kB p65/RELA Antibody Picoband® catalog # A00284-1. Tested in ELISA, Flow Cytometry, IF, 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
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-NF-kB p65/RELA Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00284-1)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived human NF-kB p65 recombinant protein (Position: F99-S551).
*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
A00284-1 is reactive to RELA in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
65-70 kDa
Calculated molecular weight
68762 MW
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
A00284-1 is guaranteed for ELISA, Flow Cytometry, IF, 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, 0.1-0.5 μg/ml
Immunocytochemistry/Immunofluorescence, 5 μg/ml
Immunoprecipitation, 0.5-2 μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
ELISA, 0.1-0.5 μg/ml
Positive Control
WB: human Hela whole cell lysates, human Raji whole cell lysates, human HepG2 whole cell lysates, human K562 whole cell lysates, rat PC-12 whole cell lysates, rat RH35 whole cell lysates, mouse RAW264.7 whole cell lysates, mouse HEPA1/6 whole cell lysates
ICC/IF: U2OS cell
IP: HepG2
FCM: Hela cell
Validation Images & Assay Conditions
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Figure 3. Immunoprecipitating NF-kB p65 in HepG2 whole cell lysate.
Western blot analysis of NF-kB p65 using anti-NF-kB p65 antibody (A00284-1).
Lane 1: HepG2 whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-NF-kB p65 antibody in HepG2 whole cell lysate,
Lane 3: anti-NF-kB p65 antibody (2μg) + HepG2 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-NF-kB p65 antigen affinity purified polyclonal antibody (A00284-1) at a dilution of 0.5 μg/mL and probed with a goat anti-rabbit IgG-HRP secondary antibody (Catalog # BA1054). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1196-200). A specific band was detected for NF-kB p65 at approximately 65-70 kDa. The expected band size for NF-kB p65 is at 60 kDa.
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Figure 1. Western blot analysis of NF-kB p65 using anti-NF-kB p65 antibody (A00284-1).
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 Raji whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: rat RH35 whole cell lysates,
Lane 7: mouse RAW264.7 whole cell lysates,
Lane 8: mouse HEPA1/6 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-kB p65 antigen affinity purified polyclonal antibody (Catalog # A00284-1) 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 NF-kB p65 at approximately 65-70 kDa. The expected band size for NF-kB p65 is at 60 kDa.
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Figure 2. IF analysis of NF-kB p65 using anti-NF-kB p65 antibody (A00284-1).
NF-kB p65 was detected in an immunocytochemical section of U2OS 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-NF-kB p65 Antibody (A00284-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 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 4. Flow Cytometry analysis of Hela cells using anti-NF-kB p65 antibody (A00284-1).
Overlay histogram showing Hela cells stained with A00284-1 (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-kB p65 Antibody (A00284-1, 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 RELA (Source: Uniprot.org, NCBI)
Gene Name
RELA
Full Name
Transcription factor p65
Weight
68762 MW
Alternative Names
Transcription factor p65; Nuclear factor NF-kappa-B p65 subunit; Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3; RELA; NFKB3 RELA CMCU, NFKB3, p65 RELA proto-oncogene, NF-kB subunit transcription factor p65|NF-kappa-B p65delta3|NF-kappa-B transcription factor p65|nuclear factor NF-kappa-B p65 subunit|nuclear factor of kappa light polypeptide gene enhancer in B-cells 3|v-rel avian reticuloendotheliosis viral oncogene homolog A
*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 RELA, check out the RELA 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 RELA: 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-NF-kB p65/RELA Antibody Picoband® (A00284-1)
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Customer Q&As
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1 Customer Q&As for Anti-NF-kB p65/RELA Antibody Picoband®
Question
Can A00284-1 be used in both membrane type pvdf and nitrocellulose?
Verified customer
Asked: 2021-02-03
Answer
The Anti-NF-KB P65/RELA Antibody Picoband™ (A00284-1) is okay to use for pvdf or nitrocellulose membrane.
Boster Scientific Support
Answered: 2021-02-03
