Product Info Summary
SKU: | M00195-1 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Mouse |
Application: | Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-ITCH/AIP4 Antibody Picoband® (monoclonal, 5E12)
SKU/Catalog Number
M00195-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ITCH/AIP4 Antibody Picoband® (monoclonal, 5E12) catalog # M00195-1. Tested in FCM, IHC, 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
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 freezing and thawing.
Cite This Product
Anti-ITCH/AIP4 Antibody Picoband® (monoclonal, 5E12) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00195-1)
Host
Mouse
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Monoclonal
Clone Number
5E12
Isotype
Mouse IgG2b
Immunogen
E.coli-derived human ITCH/AIP4 recombinant protein (Position: K17-E358).
*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
M00195-1 is reactive to ITCH in Human, Mouse, Rat
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
Observed Molecular Weight
103 kDa
Calculated molecular weight
46659 MW
Background of ITCH/AIP4
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.
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
M00195-1 is guaranteed for Flow Cytometry, 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.
Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x6 cells, Human
Positive Control
WB: human HepG2 whole cell, human 293T whole cell, human PC-3 whole cell, rat brain tissue, mouse brain tissue
IHC: human renal clear cell carcinoma tissue, human breast cancer tissue, human liver cancer tissue
FCM: U937 cell
Validation Images & Assay Conditions
![m00195 1 itch primary antibodies wb testing 1 m00195 1 itch primary antibodies wb testing 1](https://www.bosterbio.com/media/catalog/product/m/0/m00195-1-itch-primary-antibodies-wb-testing-1.jpg)
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Figure 1. Western blot analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (M00195-1).
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 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 (Catalog # M00195-1) 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-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 (Catalog # EK1001) 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.
![m00195 1 itch primary antibodies ihc testing 2 m00195 1 itch primary antibodies ihc testing 2](https://www.bosterbio.com/media/catalog/product/m/0/m00195-1-itch-primary-antibodies-ihc-testing-2.jpg)
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Figure 2. IHC analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (M00195-1).
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 μg/ml mouse anti-ITCH/AIP4 Antibody (M00195-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
![m00195 1 itch primary antibodies ihc testing 3 m00195 1 itch primary antibodies ihc testing 3](https://www.bosterbio.com/media/catalog/product/m/0/m00195-1-itch-primary-antibodies-ihc-testing-3.jpg)
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Figure 3. IHC analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (M00195-1).
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 μg/ml mouse anti-ITCH/AIP4 Antibody (M00195-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
![m00195 1 itch primary antibodies ihc testing 4 m00195 1 itch primary antibodies ihc testing 4](https://www.bosterbio.com/media/catalog/product/m/0/m00195-1-itch-primary-antibodies-ihc-testing-4.jpg)
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Figure 4. IHC analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (M00195-1).
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 μg/ml mouse anti-ITCH/AIP4 Antibody (M00195-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
![m00195 1 itch primary antibodies fcm testing 5 m00195 1 itch primary antibodies fcm testing 5](https://www.bosterbio.com/media/catalog/product/m/0/m00195-1-itch-primary-antibodies-fcm-testing-5.png)
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Figure 5. Flow Cytometry analysis of U937 cells using anti-ITCH/AIP4 antibody (M00195-1).
Overlay histogram showing U937 cells stained with M00195-1 (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 mouse anti-ITCH/AIP4 Antibody (M00195-1, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse 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 ITCH (Source: Uniprot.org, NCBI)
Gene Name
ITCH
Full Name
E3 ubiquitin-protein ligase Itchy
Weight
46659 MW
Alternative Names
Serine/threonine-protein kinase PLK1; Polo-like kinase 1; PLK-1; Serine/threonine-protein kinase 13; STPK13; PLK1; PLK Itch|6720481N21Rik, 8030492O04Rik, A, A130065M08, AIP4, C230047C07Rik|itchy, E3 ubiquitin protein ligase|E3 ubiquitin-protein ligase Itchy|HECT-type E3 ubiquitin transferase Itchy homolog
*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 ITCH, check out the ITCH Infographic
![ITCH infographic](/media/images/gene-infographic-example.jpg)
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for ITCH: 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-ITCH/AIP4 Antibody Picoband® (monoclonal, 5E12) (M00195-1)
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