Product Info Summary
| SKU: | PB9181 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-FHIT Antibody Picoband®
SKU/Catalog Number
PB9181
PB0137 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-FHIT Antibody Picoband® catalog # PB9181. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, 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
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-FHIT Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9181)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human FHIT recombinant protein (Position: M1-Q147). Human FHIT shares 90% and 87% amino acid (aa) sequences identity with mouse and rat FHIT, respectively.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9181 is reactive to FHIT in Human, Rat
Observed Molecular Weight
17 kDa
Calculated molecular weight
16.9 kDa
Background of FHIT
Bis (5'-adenosyl)-triphosphatase, also known as fragile histidine triad protein (FHIT) is an enzyme that in humans is encoded by the FHIT gene. This gene, a member of the histidine triad gene family, encodes a diadenosine P1,P3-bis (5'-adenosyl)-triphosphate adenylohydrolase involved in purine metabolism. The gene encompasses the common fragile site FRA3B on chromosome 3p14.2, where carcinogen-induced damage can lead to translocations and aberrant transcripts of this gene. In fact, aberrant transcripts from this gene have been found in about half of all esophageal, stomach, and colon carcinomas. Furthermore, FHIT has been shown to synergize with VHL, another tumor suppressor, in protecting against chemically - induced lung cancer. It also acts as a tumor suppressor of HER2/neu driven breast cancer.
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
PB9181 is guaranteed for Flow Cytometry, IF, IHC, ICC, 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, 0.1-0.5μg/ml, Human, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human Raji whole cell, rat brain tissue, rat kidney tissue
IHC: human mammary cancer tissue
ICC/IF: U20S cell
FCM: U20S cell
Validation Images & Assay Conditions
Click image to see more details
Western blot analysis of FHIT using anti-FHIT antibody (PB9181).
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 Raji whole cell lysates,
Lane 2: rat brain tissue lysates,
Lane 3: rat kidney 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 rabbit anti-FHIT antigen affinity purified polyclonal antibody (Catalog # PB9181) 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 FHIT at approximately 17 kDa. The expected band size for FHIT is at 17 kDa.
Click image to see more details
IHC analysis of FHIT using anti-FHIT antibody (PB9181).
FHIT was detected in a paraffin-embedded section of human mammary 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 1 μg/ml rabbit anti-FHIT Antibody (PB9181) 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
Flow Cytometry analysis of U20S cells using anti-FHIT antibody (PB9181).
Overlay histogram showing U20S cells stained with PB9181 (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-FHIT Antibody (PB9181,1μg/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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Click image to see more details
IF analysis of FHIT using anti-FHIT antibody (PB9181).
FHIT was detected in immunocytochemical section of U20S 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-FHIT Antibody (PB9181) 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.
Specific Publications For Anti-FHIT Antibody Picoband® (PB9181)
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1 Customer Q&As for Anti-FHIT Antibody Picoband®
Question
We are currently using anti-FHIT antibody PB9181 for human tissue, and we are happy with the IHC results. The species of reactivity given in the datasheet says human. Is it true that the antibody can work on goat tissues as well?
Verified Customer
Verified customer
Asked: 2017-07-25
Answer
The anti-FHIT antibody (PB9181) has not been tested for cross reactivity specifically with goat tissues, but 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 goat you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2017-07-25


