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product-image-AAA126863_FACS8.png FCM/FACS (Flow Cytometry) (Figure 5. Flow Cytometry analysis of U937 cells using anti-ITCH/AIP4 antibody (AAA126863).Overlay histogram showing U937 cells stained with AAA126863 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ITCH/AIP4 Antibody (AAA126863, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

Mouse ITCH/AIP4 Monoclonal Antibody | anti-ITCH antibody

Anti-ITCH/AIP4 Antibody Picoband (monoclonal, 5E12)

Gene Names
ITCH; AIF4; AIP4; NAPP1; dJ468O1.1
Reactivity
Human, Mouse, Rat
Applications
Flow Cytometry, Functional Assay, Immunohistochemistry, Western Blot
Purity
Immunogen affinity purified.
Synonyms
ITCH/AIP4, Antibody; Anti-ITCH/AIP4 Antibody Picoband (monoclonal, 5E12); anti-ITCH antibody
Ordering
Host
Mouse
Reactivity
Human, Mouse, Rat
Clonality
Monoclonal
Isotype
Mouse IgG2b
Clone Number
5E12
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 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-ITCH antibody
FCM/FACS (Flow Cytometry), IHC (Immunohistochemistry), WB (Western Blot)
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 ug/ml.
Immunogen
E Coli-derived human ITCH/AIP4 recombinant protein (Position: K17-E358).
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/FACS (Flow Cytometry)

(Figure 5. Flow Cytometry analysis of U937 cells using anti-ITCH/AIP4 antibody (AAA126863).Overlay histogram showing U937 cells stained with AAA126863 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ITCH/AIP4 Antibody (AAA126863, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

product-image-AAA126863_FACS8.png FCM/FACS (Flow Cytometry) (Figure 5. Flow Cytometry analysis of U937 cells using anti-ITCH/AIP4 antibody (AAA126863).Overlay histogram showing U937 cells stained with AAA126863 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-ITCH/AIP4 Antibody (AAA126863, 1 ug/1x10^6 cells) for 30 min at 20 degree C. DyLight488 conjugated goat anti-mouse IgG was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1 ug/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.)

IHC (Immunohistochemistry)

(Figure 4. IHC analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (AAA126863).ITCH/AIP4 was detected in a paraffin-embedded section of human liver 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 2 ug/ml mouse anti-ITCH/AIP4 Antibody (AAA126863) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

product-image-AAA126863_IHC10.jpg IHC (Immunohistochemistry) (Figure 4. IHC analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (AAA126863).ITCH/AIP4 was detected in a paraffin-embedded section of human liver 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 2 ug/ml mouse anti-ITCH/AIP4 Antibody (AAA126863) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

IHC (Immunohistochemisry)

(Figure 3. IHC analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (AAA126863).ITCH/AIP4 was detected in a paraffin-embedded section of human breast 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 2 ug/ml mouse anti-ITCH/AIP4 Antibody (AAA126863) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

product-image-AAA126863_IHC11.jpg IHC (Immunohistochemisry) (Figure 3. IHC analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (AAA126863).ITCH/AIP4 was detected in a paraffin-embedded section of human breast 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 2 ug/ml mouse anti-ITCH/AIP4 Antibody (AAA126863) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

IHC (Immunohiostchemistry)

(Figure 2. IHC analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (AAA126863).ITCH/AIP4 was detected in a paraffin-embedded section of human renal clear cell carcinoma 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 mouse anti-ITCH/AIP4 Antibody (AAA126863) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

product-image-AAA126863_IHC13.jpg IHC (Immunohiostchemistry) (Figure 2. IHC analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (AAA126863).ITCH/AIP4 was detected in a paraffin-embedded section of human renal clear cell carcinoma 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 mouse anti-ITCH/AIP4 Antibody (AAA126863) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (#SA1021) with DAB as the chromogen.)

WB (Western Blot)

(Figure 1. Western blot analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (AAA126863).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 HepG2 whole cell lysates,Lane 2: human 293T whole cell lysates,Lane 3: human PC-3 whole cell lysates,Lane 4: rat brain tissue lysates,Lane 5: 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 mouse anti-ITCH/AIP4 antigen affinity purified monoclonal antibody (#AAA126863) at 0.5 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-mouse 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 with Tanon 5200 system. A specific band was detected for ITCH/AIP4 at approximately 103 kDa. The expected band size for ITCH/AIP4 is at 103 kDa.)

product-image-AAA126863_WB15.jpg WB (Western Blot) (Figure 1. Western blot analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (AAA126863).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 HepG2 whole cell lysates,Lane 2: human 293T whole cell lysates,Lane 3: human PC-3 whole cell lysates,Lane 4: rat brain tissue lysates,Lane 5: 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 mouse anti-ITCH/AIP4 antigen affinity purified monoclonal antibody (#AAA126863) at 0.5 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-mouse 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 with Tanon 5200 system. A specific band was detected for ITCH/AIP4 at approximately 103 kDa. The expected band size for ITCH/AIP4 is at 103 kDa.)
Related Product Information for anti-ITCH antibody
ITCH is an ubiquitin-conjugating enzyme. 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 plays a role in multiple cellular processes including erythroid and lymphoid cell differentiation and the regulation of immune responses. Mutations in this gene are a cause of syndromic multisystem autoimmune disease. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene.
Product Categories/Family for anti-ITCH antibody

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
Molecular Weight
102,803 Da
NCBI Official Full Name
E3 ubiquitin-protein ligase Itchy homolog isoform 1
NCBI Official Synonym Full Names
itchy E3 ubiquitin protein ligase
NCBI Official Symbol
ITCH
NCBI Official Synonym Symbols
AIF4; AIP4; NAPP1; dJ468O1.1
NCBI Protein Information
E3 ubiquitin-protein ligase Itchy homolog; NFE2-associated polypeptide 1; atrophin-1 interacting protein 4; itchy E3 ubiquitin protein ligase homolog; dJ468O1.1 (atrophin 1 interacting protein 4 (AIP4))
UniProt Protein Name
E3 ubiquitin-protein ligase Itchy homolog
UniProt Gene Name
ITCH
UniProt Synonym Gene Names
Itch; AIP4; NAPP1
UniProt Entry Name
ITCH_HUMAN

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Product Notes

The ITCH itch (Catalog #AAA126863) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-ITCH/AIP4 Antibody Picoband (monoclonal, 5E12) reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's ITCH/AIP4 can be used in a range of immunoassay formats including, but not limited to, FCM/FACS (Flow Cytometry), IHC (Immunohistochemistry), WB (Western Blot). Researchers should empirically determine the suitability of the ITCH itch 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 "ITCH/AIP4, Monoclonal 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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