Rabbit IgG polyclonal antibody for Indoleamine 2,3-dioxygenase 2(IDO2) detection. Tested with WB, FCM in Human.
|Applications||Flow Cytometry, WB|
|Product Name||Anti-INDOL1/IDO2 Antibody
See all IDO2 primary antibodies, ELISA kits and proteins
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Polyclonal antibody for IDO2 detection. Host: Rabbit.Size: 100μg/vial. Tested applications: WB, FCM. Reactive species: Human. IDO2 information: Molecular Weight: 45424 MW; Tissue Specificity: Detected in liver, small intestine, spleen, placenta, thymus, lung, brain, kidney, and colon.|
|Cite This Product||Anti-INDOL1/IDO2 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1612)|
|Specificity||Anti-INDOL1/IDO2 Antibody (PA1612) reacts with Human IDO2, in native form and recombinant. Superfamily members of IDO2 are not reactive to PA1612.|
|Contents/Buffer||Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.|
|Reconstitution||Add 0.2ml of distilled water will yield a concentration of 500ug/ml.|
|Immunogen||A synthetic peptide corresponding to a sequence at the N-terminus of human INDOL1(1-20aa MLHFHYYDTSNKIMEPHRPN).|
Our Boster Quality Guarantee for Anti-INDOL1/IDO2 Antibody covers its use in the following applications.
*The recommended dilution ratios/concentrations are for reference only and optimal dilutions/concentrations should be determined by the end user.
Assay Dilutions Overview
Western blot, 0.1-0.5μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
Boster's Compatible Products
The following reagents are used to generate the images below for Anti-INDOL1/IDO2 Antibody (PA1612).Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot.
Images And Assay Conditions
Figure 2. Flow Cytometry analysis of SiHa cells using anti-IDO2 antibody (PA1612).
Overlay histogram showing SiHa cells stained with PA1612 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-IDO2 Antibody (PA1612, 1Î¼g/1x106 cells) for 30 min at 20Â°C. DyLightÂ®488 conjugated goat anti-rabbit IgG (BA1127, 5-10Î¼g/1x106 cells) was used as secondary antibody for 30 minutes at 20Â°C. Isotype control antibody (Green line) was rabbit IgG (1Î¼g/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Figure 1. Western blot analysis of IDO2 using anti- IDO2 antibody (PA1612).
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 A549 whole cell lysates (positive control),
Lane 2: human placenta tissue lysates (positive control),
Lane 3: human Caco-2 whole cell lysates (positive control),
Lane 4: human PC-3 whole cell lysates (negative control).
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- IDO2 antigen affinity purified polyclonal antibody (Catalog # PA1612) at 0.5 Î¼g/mL overnight at 4Â°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 IDO2 at approximately 47KD. The expected band size for IDO2 is at 47KD.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Indoleamine 2,3-dioxygenase 2|
|Tissue Specificity||Detected in liver, small intestine, spleen, placenta, thymus, lung, brain, kidney, and colon. .|
|Alternative Names||Indoleamine 2,3-dioxygenase 2;IDO-2;1.13.11.-;Indoleamine 2,3-dioxygenase-like protein 1;Indoleamine-pyrrole 2,3-dioxygenase-like protein 1;IDO2;INDOL1;|
|Molecular Weight||45424 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Catalyzes the first and rate-limiting step in the kynurenine pathway of tryptophan catabolism. .|
*You can search these to find other products in these research areas.
|Background||IDO2(Indoleamine 2,3-dioxygenase 2), also called INDOLEAMINE 2,3-DIOXYGENASE-LIKE 1 or INDOL1, is an enzyme encoded by the INDOL1 gene which metabolizes tryptophan in the kynurenine pathway. By genomic sequence analysis, the INDOL1 gene is mapped on chromosome 8p12 just downstream of the INDO gene. And its exact cytogenetic location is 8p11.21. By database analysis using INDO as probe, followed by RT-PCR of total RNA from various tissues, IDO2 is cloned by human and mouse INDOL1. INDOL1 catabolizes tryptophan as determined by Kyn production, but unlike INDO, is inhibited by D-1-methyl-tryptophan(D-1MT) but not the L-1MT stereoisomer. The Gene Structure of the INDOL1 has 11 exons and spans 74 kb.|
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Guaranteed product quality
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Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugationA: Yes, please contact us at [email protected] for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WBA: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugationA: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact [email protected]
Q: Does this antibody only recognize IDO2? Does this antibody have cross-reactivity with IDO1?A: This antibody has no cross reactivity with IDO1 (We got negative result in WB).
Q: What should I use for negative control?A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signalA: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?A: You can find the immunogen sequence under "
Q: What is the expected band size? Why is it different than the observed band size?A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:<br>1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?A: Check our protocols under the tech support tab.