Product Info Summary
SKU: | A00195-1 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-ITCH/AIP4 Antibody Picoband™
SKU/Catalog Number
A00195-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ITCH/AIP4 Antibody Picoband™ catalog # A00195-1. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
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-ITCH/AIP4 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00195-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
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
A00195-1 is reactive to Itch in Human, Mouse, Rat
Applications
A00195-1 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
103 kDa
Calculated molecular weight
98.994kDa
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.
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Assay dilution & Images
Reconsitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
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, 0.1-0.25μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of ITCH using anti-ITCH antibody (A00195-1).
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 K562 whole cell lysates,
Lane 2: human Raji whole cell lysates,
Lane 3: rat C6 whole cell lysates,
Lane 4: mouse NIH/3T3 whole cell lysates,
Lane 5: human HepG2 whole cell lysates,
Lane 6: human A549 whole cell lysates,
Lane 7: human THP-1 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-ITCH antigen affinity purified polyclonal antibody (Catalog # A00195-1) at 0.25 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 ITCH at approximately 103KD. The expected band size for ITCH is at 103KD.
Click image to see more details
Figure 2. IHC analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (A00195-1).
ITCH/AIP4 was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ITCH/AIP4 Antibody (A00195-1) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (A00195-1).
ITCH/AIP4 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-ITCH/AIP4 Antibody (A00195-1) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IF analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (A00195-1).
ITCH/AIP4 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-ITCH/AIP4 Antibody (A00195-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 5. Flow Cytometry analysis of SiHa cells using anti-ITCH/AIP4 antibody (A00195-1).
Overlay histogram showing SiHa cells stained with A00195-1 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ITCH/AIP4 Antibody (A00195-1,1μg/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 Itch (Source: Uniprot.org, NCBI)
Gene Name
Itch
Full Name
E3 ubiquitin-protein ligase Itchy
Weight
98.994kDa
Alternative Names
AIF4; AIP4; AIP4dJ468O1.1 (atrophin 1 interacting protein 4 (AIP4)); atrophin-1 interacting protein 4; Atrophin-1-interacting protein 4; dJ468O1.1; EC 6.3.2; EC 6.3.2.-; ITCH; itchy (mouse homolog) E3 ubiquitin protein ligase; itchy E3 ubiquitin protein ligase homolog (mouse); itchy homolog E3 ubiquitin protein ligase; NAPP1; NAPP1E3 ubiquitin-protein ligase Itchy homolog; NFE2-associated polypeptide 1; ubiquitin protein ligase ITCH 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
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™ (A00195-1)
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No publications found for A00195-1
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