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Product Info Summary
| SKU: | A01595-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, ICC, WB |
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Product info
Product Name
Anti-NEDD4-2/NEDD4L Antibody Picoband®
SKU/Catalog Number
A01595-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-NEDD4-2/NEDD4L Antibody Picoband® catalog # A01595-1. Tested in ELISA, Flow Cytometry, 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
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-NEDD4-2/NEDD4L Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01595-1)
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 human NEDD4-2 recombinant protein (Position: N703-D975).
*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
A01595-1 is reactive to NEDD4L in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
130 kDa
Calculated molecular weight
111.932kDa
Background of NEDD4L
Neural precursor cell expressed developmentally downregulated gene 4-like (NEDD4L) or NEDD4-2 (NEDD4-2) is an enzyme (ubiquitin ligase) of the NEDD4 family. This gene encodes a member of the Nedd4 family of HECT domain E3 ubiquitin ligases. HECT domain E3 ubiquitin ligases transfer ubiquitin from E2 ubiquitin-conjugating enzymes to protein substrates, thus targeting specific proteins for lysosomal degradation. The encoded protein mediates the ubiquitination of multiple target substrates and plays a critical role in epithelial sodium transport by regulating the cell surface expression of the epithelial sodium channel, ENaC. Single nucleotide polymorphisms in this gene may be associated with essential hypertension. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene.
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
A01595-1 is guaranteed for ELISA, Flow Cytometry, 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, 0.1-0.5μg/ml
Immunohistochemistry (Frozen Section), 0.5-1μg/ml
Immunocytochemistry, 0.5-1μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
ELISA, 0.1-0.5μg/ml
Positive Control
WB: human 22RV1 whole cell, human Jurkat whole cell, rat pancreas tissue, mouse pancreas tissue, mouse NIH3T3 whole cell
FCM: U20S cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of NEDD4-2 using anti-NEDD4-2 antibody (A01595-1).
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 50ug of sample under reducing conditions.
Lane 1: human 22RV1 whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: rat pancreas tissue lysates,
Lane 4: mouse pancreas tissue lysates,
Lane 5: mouse NIH3T3 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-NEDD4-2 antigen affinity purified polyclonal antibody (Catalog # A01595-1) 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 1:10000 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 NEDD4-2 at approximately 130KD. The expected band size for NEDD4-2 is at 112KD.
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Figure 2. Flow Cytometry analysis of U20S cells using anti-NEDD4-2 antibody (A01595-1).
Overlay histogram showing U20S cells stained with A01595-1 (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-NEDD4-2 Antibody (A01595-1,1μg/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 (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For NEDD4L (Source: Uniprot.org, NCBI)
Gene Name
NEDD4L
Full Name
E3 ubiquitin-protein ligase NEDD4-like
Weight
111.932kDa
Alternative Names
E3 ubiquitin-protein ligase NEDD4-like; HECT-type E3 ubiquitin transferase NED4L; NEDD4.2; Nedd4-2; NEDD4L; KIAA0439; NEDL3 NEDD4L NEDD4-2, NEDD4.2, PVNH7, RSP5, hNEDD4-2 NEDD4 like E3 ubiquitin protein ligase E3 ubiquitin-protein ligase NEDD4-like|HECT-type E3 ubiquitin transferase NED4L|neural precursor cell expressed, developmentally down-regulated 4-like, E3 ubiquitin protein ligase|ubiquitin-protein ligase Rsp5
*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 NEDD4L, check out the NEDD4L 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 NEDD4L: 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-NEDD4-2/NEDD4L Antibody Picoband® (A01595-1)
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Customer Q&As
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6 Customer Q&As for Anti-NEDD4-2/NEDD4L Antibody Picoband®
Question
I am interested in to test anti-NEDD4-2/NEDD4L antibody A01595-1 on mouse prostate for research purposes, then I may be interested in using anti-NEDD4-2/NEDD4L antibody A01595-1 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
J. Gonzalez
Verified customer
Asked: 2020-03-13
Answer
The products we sell, including anti-NEDD4-2/NEDD4L antibody A01595-1, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2020-03-13
Question
Can you help my question with product A01595-1, anti-NEDD4-2/NEDD4L antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
Verified Customer
Verified customer
Asked: 2019-12-16
Answer
We suggest not storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free A01595-1 anti-NEDD4-2/NEDD4L antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2019-12-16
Question
We appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for prostate using anti-NEDD4-2/NEDD4L antibody A01595-1. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-09-19
Answer
We appreciate the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2019-09-19
Question
Is there a BSA free version of anti-NEDD4-2/NEDD4L antibody A01595-1 available?
Verified Customer
Verified customer
Asked: 2019-08-15
Answer
We appreciate your recent telephone inquiry. I can confirm that some lots of this anti-NEDD4-2/NEDD4L antibody A01595-1 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2019-08-15
Question
I see that the anti-NEDD4-2/NEDD4L antibody A01595-1 works with WB, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2018-07-31
Answer
You can find protocols for WB on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to [email protected]
Boster Scientific Support
Answered: 2018-07-31
Question
We are currently using anti-NEDD4-2/NEDD4L antibody A01595-1 for rat tissue, and we are satisfied with the IHC results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on bovine tissues as well?
P. Brown
Verified customer
Asked: 2013-11-29
Answer
The anti-NEDD4-2/NEDD4L antibody (A01595-1) has not been validated for cross reactivity specifically with bovine 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 bovine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2013-11-29
