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ITCH antibody (M00195-1) performs well in Flow Cytometry, IHC, WB and shows reactivity with Human, Mouse, Rat.
- Validated for inter-lot consistency, specificity, and reproducibility
- Committed to quality since 1993
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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.
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.
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Anti-ITCH/AIP4 Antibody Picoband® (monoclonal, 5E12)
ITCH antibody
| Reactive Species | Host | Application |
|---|---|---|
| Human, Mouse, Rat | Mouse | Flow Cytometry, IHC, WB |
Tested Applications
ITCH antibody was positively detected by WB in:
human HepG2 whole cell, human 293T whole cell, human PC-3 whole cell, rat brain tissue, mouse brain tissue
Suggested blocking solution with 5% non-fat milk or BSA; (*)Recommended protein loading: 20-40 ug per lane
ITCH antibody was positively detected by IHC in:
human renal clear cell carcinoma tissue, human breast cancer tissue, human liver cancer tissue
Use TE buffer pH 9.0 for antigen retrieval; (*) citrate buffer pH 6.0 is an alternative.
ITCH antibody was positively detected by FC in:
U937 cell
Recommend Dilution
Application
Dilution
Species
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
* It is recommended to optimize the amount of this reagent in each experimental system to achieve the best results.
Note: For flow cytometry (FCM), validated for fixed cells only. Not validated for FACS or surface flow cytometry using live cells.
Reactive Data
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WB
IHC
IF
Human
Tested
Tested
Recommend
Rat
Tested
Mouse
Tested
Product Information
Product Name
Anti-ITCH/AIP4 Antibody Picoband® (monoclonal, 5E12)
View all ITCH Antibodies
View all ITCH Antibodies
Form
Lyophilized
Uniprot
Gene ID
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.
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).
Cross-Reactivity
No cross-reactivity with other proteins.
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
Observed Molecular Weight
103 kDa
Calculated Molecular Weight
102.8 kDa
Background of ITCH
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.
Cite This Product
Anti-ITCH/AIP4 Antibody Picoband® (monoclonal, 5E12) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00195-1)
Antibody Homology Across Species
The results generated by this homology analysis tool for commonly used animal species are shown below. For unlisted species, please refer to the homology analysis between the immunogen sequence and the corresponding species.
A homology of ≥90% is recommended. Generally, higher homology indicates a greater potential for cross-species reactivity. However, protein expression levels may vary among different species, tissues, and cell types. Therefore, the homology analysis results are for reference only and do not guarantee antibody cross-reactivity or experimental performance.
Human
100%
High Homology
Homology ≥ 90%High Homology
70% ≤ Homology < 90%Moderate Homology
Homology < 70%Low homology
Protocols
Western Blot protocol for ITCH antibody
Immunohistochemistry (IHC) protocol for ITCH antibody
Flow Cytometry (FC) protocol for ITCH antibody
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Product Promise
We promise Anti-ITCH/AIP4 Antibody Picoband® (monoclonal, 5E12) performs excellently in relevant applications for Flow Cytometry, IHC, WB. If it doesn't perform as promised, we will refund the product. Learn more
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M00195-1
Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
*Sample sizes are prepared on demand and will take extra lead time. (cannot be conjugated)
$370.00
Troubleshooting
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We promise all of our products perform as described in datasheets.
This product is for Research Use Only. Not for use in diagnostic procedures.
Update date:
Oct 2, 2026