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FCM (Flow Cytometry) (Figure 6. Flow Cytometry analysis of HEPA1-6 cells using anti-Nedd4 antibody (AAA19414).Overlay histogram showing HEPA1-6 cells stained with AAA19414 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Nedd4 Antibody (AAA19414, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Rabbit anti-Mouse, Rat Nedd4 Polyclonal Antibody | anti-NEDD4 antibody

Anti-Nedd4 Antibody Picoband

Gene Names
Nedd4; Nedd4a; Nedd4-1; AA959633; AL023035; AU019897; mKIAA0093; E430025J12Rik
Reactivity
Mouse, Rat
Applications
Western Blot, Immunohistochemistry, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
Nedd4; Polyclonal Antibody; Anti-Nedd4 Antibody Picoband; anti-NEDD4 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Natural and recombinant Human CSF3
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Concentration
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml. (varies by lot)
Applicable Applications for anti-NEDD4 antibody
Western Blot (WB), Immunohistochemistry (IHC), Flow Cytometry (FC/FACS), ELISA (EIA)
Application Notes
WB: 0.25-0.5 ug/ml, Mouse, Rat
IHC-P: 2-5 ug/ml, Mouse, Rat
FC/FACS: 1-3 ug/1x10^6 cells, Mouse
Direct ELISA: 0.1-0.5 ug/ml, Mouse
Tested Species: In-house tested species with positive results.
Enhanced Chemiluminescent Kit with anti-Rabbit IgG for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit for IHC(P).
Samples
Cell culture supernatants, serum and plasma (heparin, EDTA, citrate)
Detection Range
31.2 pg/ml - 2,000 pg/ml
Immunogen
E Coli-derived mouse Nedd4 recombinant protein (Position: A7-L655).
Preparation and Storage
Store at -20 degree C for one year from date of receipt. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for six months. Avoid repeated freezing and thawing.

FCM (Flow Cytometry)

(Figure 6. Flow Cytometry analysis of HEPA1-6 cells using anti-Nedd4 antibody (AAA19414).Overlay histogram showing HEPA1-6 cells stained with AAA19414 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Nedd4 Antibody (AAA19414, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

FCM (Flow Cytometry) (Figure 6. Flow Cytometry analysis of HEPA1-6 cells using anti-Nedd4 antibody (AAA19414).Overlay histogram showing HEPA1-6 cells stained with AAA19414 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Nedd4 Antibody (AAA19414, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-rabbit IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IHC (Immunohistochemistry)

(Figure 5. IHC analysis of Nedd4 using anti-Nedd4 antibody (AAA19414).Nedd4 was detected in a paraffin-embedded section of rat brain 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 2 ug/ml rabbit anti-Nedd4 Antibody (AAA19414) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 5. IHC analysis of Nedd4 using anti-Nedd4 antibody (AAA19414).Nedd4 was detected in a paraffin-embedded section of rat brain 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 2 ug/ml rabbit anti-Nedd4 Antibody (AAA19414) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 4. IHC analysis of Nedd4 using anti-Nedd4 antibody (AAA19414).Nedd4 was detected in a paraffin-embedded section of rat brain 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 2 ug/ml rabbit anti-Nedd4 Antibody (AAA19414) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 4. IHC analysis of Nedd4 using anti-Nedd4 antibody (AAA19414).Nedd4 was detected in a paraffin-embedded section of rat brain 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 2 ug/ml rabbit anti-Nedd4 Antibody (AAA19414) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 3. IHC analysis of Nedd4 using anti-Nedd4 antibody (AAA19414).Nedd4 was detected in a paraffin-embedded section of mouse skeletal muscle 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 2 ug/ml rabbit anti-Nedd4 Antibody (AAA19414) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 3. IHC analysis of Nedd4 using anti-Nedd4 antibody (AAA19414).Nedd4 was detected in a paraffin-embedded section of mouse skeletal muscle 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 2 ug/ml rabbit anti-Nedd4 Antibody (AAA19414) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

IHC (Immunohistochemistry)

(Figure 2. IHC analysis of Nedd4 using anti-Nedd4 antibody (AAA19414).Nedd4 was detected in a paraffin-embedded section of mouse brain 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 2 ug/ml rabbit anti-Nedd4 Antibody (AAA19414) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

IHC (Immunohistochemistry) (Figure 2. IHC analysis of Nedd4 using anti-Nedd4 antibody (AAA19414).Nedd4 was detected in a paraffin-embedded section of mouse brain 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 2 ug/ml rabbit anti-Nedd4 Antibody (AAA19414) overnight at 4 degree C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of Nedd4 using anti-Nedd4 antibody (AAA19414).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: rat L6 whole cell lysates,Lane 2: mouse skeletal muscle tissue lysates,Lane 3: mouse lung tissue lysates,Lane 4: mouse B16-F10 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-Nedd4 antigen affinity purified polyclonal antibody (#AAA19414) at 0.25 ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for Nedd4 at approximately 115 kDa. The expected band size for Nedd4 is at 149 kDa.)

WB (Western Blot) (Figure 1. Western blot analysis of Nedd4 using anti-Nedd4 antibody (AAA19414).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: rat L6 whole cell lysates,Lane 2: mouse skeletal muscle tissue lysates,Lane 3: mouse lung tissue lysates,Lane 4: mouse B16-F10 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-Nedd4 antigen affinity purified polyclonal antibody (#AAA19414) at 0.25 ug/mL overnight at 4 degree 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 with Tanon 5200 system. A specific band was detected for Nedd4 at approximately 115 kDa. The expected band size for Nedd4 is at 149 kDa.)
Related Product Information for anti-NEDD4 antibody
Background: Granulocyte colony-stimulating factor (G-CSF) is a member of the CSF family of hormone-like glycoprotein that regulates hematopoietic cell proliferation and differentiation, and it almost exclusively stimulates the colony formation of granulocytes from committed precursor cells in semisolid agar culture. G-CSF is also termed colony stimulating factor-3, and a single gene of which codes for a 177 or 180 amino acid mature protein of molecular weight 19,600. Functionally, it specifically stimulates the proliferation and differentiation of the progenitor cells for granulocytes. The effect of G-CSF on myeloid leukemias is unique among colony stimulating factors in driving the leukemic cells from a self-renewing malignant state to a mature differentiated phenotype with the concomitant loss of tumorigenicity. Besides, it also prevents cardiac remodeling after myocardial infarction by activating the Jak-Stat pathway in cardiomyocytes. The recombinant form of hG-CSF is capable of supporting neutrophil proliferation in a CFU-GM assay as well as early erythroid colonies and mixed colony formation. Human gene coding for G-CSF is assigned to the q21-q22 region of chromosome 17. The standard product used in this kit is recombinant human G-CSF, consisting of 175 amino acids with the molecular mass of 18.8KDa.
Product Categories/Family for anti-NEDD4 antibody

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
UniProt Accession #
Molecular Weight
102,706 Da
NCBI Official Full Name
E3 ubiquitin-protein ligase NEDD4
NCBI Official Synonym Full Names
neural precursor cell expressed, developmentally down-regulated 4
NCBI Official Symbol
Nedd4
NCBI Official Synonym Symbols
Nedd4a; Nedd4-1; AA959633; AL023035; AU019897; mKIAA0093; E430025J12Rik
NCBI Protein Information
E3 ubiquitin-protein ligase NEDD4; NEDD-4; neural precursor cell expressed, developmentally down-regulted gene 4; neural precursor cell expressed, developmentally down-regulated gene 4; neural precursor cell expressed, developmentally down-regulated gene
UniProt Protein Name
E3 ubiquitin-protein ligase NEDD4
UniProt Gene Name
Nedd4
UniProt Synonym Gene Names
Kiaa0093; Nedd-4; Nedd4-1; Nedd4a; NEDD-4
UniProt Entry Name
NEDD4_MOUSE

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Product Notes

The NEDD4 nedd4 (Catalog #AAA19414) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-Nedd4 Antibody Picoband reacts with Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's Nedd4 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC), Flow Cytometry (FC/FACS), ELISA (EIA). WB: 0.25-0.5 ug/ml, Mouse, Rat IHC-P: 2-5 ug/ml, Mouse, Rat FC/FACS: 1-3 ug/1x10^6 cells, Mouse Direct ELISA: 0.1-0.5 ug/ml, Mouse Tested Species: In-house tested species with positive results. Enhanced Chemiluminescent Kit with anti-Rabbit IgG for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit for IHC(P). Researchers should empirically determine the suitability of the NEDD4 nedd4 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Nedd4, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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