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Product Info Summary
| SKU: | M00984 |
|---|---|
| Size: | 100 μl |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IP, IHC, WB |
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Product info
Product Name
Anti-NEDD4 Rabbit Monoclonal Antibody
SKU/Catalog Number
M00984
Size
100 μl
Form
Liquid
Description
Boster Bio Anti-NEDD4 Rabbit Monoclonal Antibody catalog # M00984. Tested in WB, IHC, IP, Flow Cytometry applications. This antibody reacts with Human, Mouse, Rat.
Storage & Handling
Store at -20°C for one year. For short term storage and frequent use, store at 4°C for up to one month. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-NEDD4 Rabbit Monoclonal Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00984)
Host
Rabbit
Contents
Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol, 0.4-0.5mg/ml BSA.
Clonality
Monoclonal
Clone Number
AEGB-14
Isotype
Rabbit IgG
Immunogen
A synthesized peptide derived from human NEDD4
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Reactive Species
M00984 is reactive to NEDD4 in Human, Mouse, Rat
Reconstitution
Restore with deionized water (or equivalent) for reconstitution volume of 1.0 mL
Observed Molecular Weight
135 kDa
Calculated molecular weight
105-150 kDa
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
M00984 is guaranteed for Flow Cytometry, IP, 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.
WB 1:1000-1:5000
IHC 1:50-1:200
IP 1:50
FC 1:50
Positive Control
WB: human A549 whole cell, human SH-SY5Y whole cell, human U251 whole cell, human 293T whole cell, rat brain tissue, mouse brain tissue
IHC: human bladder cancer tissue, human pancreas cancer tissue
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of NEDD4 using anti-NEDD4 antibody (M00984).
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 A549 whole cell lysates,
Lane 2: human SH-SY5Y whole cell lysates,
Lane 3: human U251 whole cell lysates,
Lane 4: human 293T whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: mouse brain 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-NEDD4 antigen affinity purified monoclonal antibody (Catalog # M00984) at 1:1000 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 1:500 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 at approximately 135 kDa. The expected band size for NEDD4 is at 105-150 kDa.
Click image to see more details
Figure 2. IHC analysis of NEDD4 using anti-NEDD4 antibody (M00984).
NEDD4 was detected in a paraffin-embedded section of human bladder 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 1:50 rabbit anti-NEDD4 Antibody (M00984) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of NEDD4 using anti-NEDD4 antibody (M00984).
NEDD4 was detected in a paraffin-embedded section of human pancreas 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 1:50 rabbit anti-NEDD4 Antibody (M00984) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Protein Target Info & Infographic
Gene/Protein Information For NEDD4 (Source: Uniprot.org, NCBI)
Gene Name
NEDD4
Full Name
E3 ubiquitin-protein ligase NEDD4
Weight
105-150 kDa
Alternative Names
Protein S100-B;S-100 protein beta chain;S-100 protein subunit beta;S100 calcium-binding protein B;S100B; NEDD4 NEDD4-1, RPF1 NEDD4 E3 ubiquitin protein ligase E3 ubiquitin-protein ligase NEDD4|HECT-type E3 ubiquitin transferase NEDD4|cell proliferation-inducing gene 53 protein|neural precursor cell expressed, developmentally down-regulated 4, E3 ubiquitin protein ligase|receptor-potentiating factor 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 NEDD4, check out the NEDD4 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 NEDD4: 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 Rabbit Monoclonal Antibody (M00984)
Hello CJ!
M00984 has been cited in 1 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
The Novel Dual GLP-1/GIP Receptor Agonist DA-CH5 Is Superior to Single GLP-1 Receptor Agonists in the MPTP Model of Parkinson's Disease Lingyu Zhang J Parkinson's Dis. 2020;10(2):523-542. DOI: 10.3233/JPD-191768.
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Customer Q&As
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15 Customer Q&As for Anti-NEDD4 Rabbit Monoclonal Antibody
Question
Do you have a BSA free version of anti-NEDD4 Monoclonal antibody M00984 available?
Verified Customer
Verified customer
Asked: 2020-05-07
Answer
Thank you for your recent telephone inquiry. I can confirm that some lots of this anti-NEDD4 Monoclonal antibody M00984 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: 2020-05-07
Question
Is this M00984 anti-NEDD4 Monoclonal antibody reactive to the isotypes of NEDD4?
Verified Customer
Verified customer
Asked: 2020-05-05
Answer
The immunogen of M00984 anti-NEDD4 Monoclonal antibody is A synthesized peptide derived from human NEDD4 E3 ubiquitin-protein ligase which accepts ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfers the ubiquitin to targeted substrates. Involved in the pathway leading to the degradation of VEGFR-2/KDFR, independently of its ubiquitin-ligase activity. Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2020-05-05
Question
I am looking for to test anti-NEDD4 Monoclonal antibody M00984 on mouse trachea for research purposes, then I may be interested in using anti-NEDD4 Monoclonal antibody M00984 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
B. Krishna
Verified customer
Asked: 2020-02-25
Answer
The products we sell, including anti-NEDD4 Monoclonal antibody M00984, 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: 2020-02-25
Question
Can you help my question with product M00984, anti-NEDD4 Monoclonal 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: 2020-01-14
Answer
We do not advise 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 M00984 anti-NEDD4 Monoclonal 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: 2020-01-14
Question
Our lab were content with the WB result of your anti-NEDD4 Monoclonal antibody. However we have observed positive staining in biceps brachii cytoplasm. cell membrane using this antibody. Is that expected? Could you tell me where is NEDD4 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2019-12-25
Answer
From literature, biceps brachii does express NEDD4. Generally NEDD4 expresses in cytoplasm. cell membrane. Regarding which tissues have NEDD4 expression, here are a few articles citing expression in various tissues:
Adipose tissue, Pubmed ID: 17974005
Bone marrow, Pubmed ID: 7788527
Brain, and Testis, Pubmed ID: 15489334
Cervix carcinoma, Pubmed ID: 18669648, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Leukemic T-cell, Pubmed ID: 19690332
Trachea, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2019-12-25
Question
We ordered your anti-NEDD4 Monoclonal antibody for WB on biceps brachii a few years ago. I am using rat, and We intend to use the antibody for IP next. We are interested in examining biceps brachii as well as bone marrow in our next experiment. Could you please give me some suggestion on which antibody would work the best for IP?
Verified Customer
Verified customer
Asked: 2019-11-19
Answer
I viewed the website and datasheets of our anti-NEDD4 Monoclonal antibody and it seems that M00984 has been tested on rat in both WB and IP. Thus M00984 should work for your application. Our Boster satisfaction guarantee will cover this product for IP in rat even if the specific tissue type has not been validated. We do have a comprehensive range of products for IP detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2019-11-19
Question
We are currently using anti-NEDD4 Monoclonal antibody M00984 for mouse tissue, and we are well pleased with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on horse tissues as well?
Verified Customer
Verified customer
Asked: 2019-07-24
Answer
The anti-NEDD4 Monoclonal antibody (M00984) has not been tested for cross reactivity specifically with horse tissues, though 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: 2019-07-24
Question
I see that the anti-NEDD4 Monoclonal antibody M00984 works with WB, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2019-07-18
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: 2019-07-18
Question
Would anti-NEDD4 Monoclonal antibody M00984 work for WB with trachea?
Verified Customer
Verified customer
Asked: 2019-06-18
Answer
According to the expression profile of trachea, NEDD4 is highly expressed in trachea. So, it is likely that anti-NEDD4 Monoclonal antibody M00984 will work for WB with trachea.
Boster Scientific Support
Answered: 2019-06-18
Question
We appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for trachea using anti-NEDD4 Monoclonal antibody M00984. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2019-06-14
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: 2019-06-14
Question
Is a blocking peptide available for product anti-NEDD4 Monoclonal antibody (M00984)?
Verified Customer
Verified customer
Asked: 2019-06-05
Answer
We do provide the blocking peptide for product anti-NEDD4 Monoclonal antibody (M00984). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2019-06-05
Question
See below the WB image, lot number and protocol we used for trachea using anti-NEDD4 Monoclonal antibody M00984. Please let me know if you require anything else.
Verified Customer
Verified customer
Asked: 2018-08-15
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2018-08-15
Question
We have seen staining in mouse leukemic t-cell. Any tips? Is anti-NEDD4 Monoclonal antibody supposed to stain leukemic t-cell positively?
Verified Customer
Verified customer
Asked: 2018-08-03
Answer
According to literature leukemic t-cell does express NEDD4. According to Uniprot.org, NEDD4 is expressed in biceps brachii, bone marrow, trachea, adipose tissue, brain testis, cervix carcinoma, leukemic t-cell, cervix carcinoma erythroleukemia, among other tissues. Regarding which tissues have NEDD4 expression, here are a few articles citing expression in various tissues:
Adipose tissue, Pubmed ID: 17974005
Bone marrow, Pubmed ID: 7788527
Brain, and Testis, Pubmed ID: 15489334
Cervix carcinoma, Pubmed ID: 18669648, 20068231
Cervix carcinoma, and Erythroleukemia, Pubmed ID: 23186163
Leukemic T-cell, Pubmed ID: 19690332
Trachea, Pubmed ID: 14702039
Boster Scientific Support
Answered: 2018-08-03
Question
I was wanting to use your anti-NEDD4 Monoclonal antibody for WB for mouse trachea on frozen tissues, but I want to know if it has been tested for this particular application. Has this antibody been tested and is this antibody a good choice for mouse trachea identification?
Verified Customer
Verified customer
Asked: 2017-05-17
Answer
You can see on the product datasheet, M00984 anti-NEDD4 Monoclonal antibody has been tested for IP, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in mouse trachea in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2017-05-17
Question
Does M00984 anti-NEDD4 Monoclonal antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
F. Huang
Verified customer
Asked: 2015-03-17
Answer
It shows on the product datasheet, M00984 anti-NEDD4 Monoclonal antibody as been validated on WB. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2015-03-17
