Product Info Summary
SKU: | PB9496 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Monkey, Mouse, Rat |
Host: | Rabbit |
Application: | Flow Cytometry, WB |
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Product info
Product Name
Anti-ITCH/AIP4 Antibody Picoband™
SKU/Catalog Number
PB9496
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ITCH/AIP4 Antibody Picoband™ catalog # PB9496. Tested in WB, Flow Cytometry applications. This antibody reacts with Human, Monkey, 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 # PB9496)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence in the middle region of human ITCH/AIP4, different from the related mouse sequence by one amino acid.
*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
PB9496 is reactive to Itch in Human, Monkey, Mouse, Rat
Applications
PB9496 is guaranteed for Flow Cytometry, WB Boster Guarantee
Observed Molecular Weight
103 kDa
Calculated molecular weight
102803 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.
Assay dilution & Images
Reconstitution
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, 0.1-0.5μg/ml, Human, Monkey, Mouse, Rat
Flow Cytometry(Fixed), 1-3 μg/1x106 cells, Human
Validation Images & Assay Conditions
![pb9496 itch primary antibodies wb testing 1 pb9496 itch primary antibodies wb testing 1](https://www.bosterbio.com/media/catalog/product/p/b/pb9496-itch-primary-antibodies-wb-testing-1.jpg)
Click image to see more details
Figure 1. Western blot analysis of ITCH/AIP4 using anti-ITCH/AIP4 antibody (PB9496).
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 K562 whole cell lysates,
Lane 2: human Raji whole cell lysates,
Lane 3: human 293T whole cell lysates,
Lane 4: monkey COS-7 whole cell lysates,
Lane 5: human HepG2 whole cell lysates,
Lane 6: human A549 whole cell lysates,
Lane 7: rat C6 whole cell lysates,
Lane 8: mouse NIH/3T3 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-ITCH/AIP4 antigen affinity purified polyclonal antibody (Catalog # PB9496) 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-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 (Catalog # EK1002) 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.
![pb9496 itch primary antibodies fcm testing 2 pb9496 itch primary antibodies fcm testing 2](https://www.bosterbio.com/media/catalog/product/p/b/pb9496-itch-primary-antibodies-fcm-testing-2.jpg)
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Figure 2. Flow Cytometry analysis of SiHa cells using anti-ITCH antibody (PB9496).
Overlay histogram showing SiHa cells stained with PB9496 (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 rabbit anti-ITCH Antibody (PB9496, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit 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
102803 MW
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
![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™ (PB9496)
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Customer Q&As
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1 Customer Q&As for Anti-ITCH/AIP4 Antibody Picoband™
Question
We are currently using anti-ITCH/AIP4 antibody PB9496 for human tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human. Is it possible that the antibody can work on bovine tissues as well?
J. Williams
Verified customer
Asked: 2016-03-04
Answer
The anti-ITCH/AIP4 antibody (PB9496) has not been tested for cross reactivity specifically with bovine 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 bovine you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2016-03-04