Anti-FHIT Antibody Picoband™

Product Name

Anti-FHIT Antibody Picoband™

View all FHIT Antibodies

SKU/Catalog Number

PB9181

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.

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 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

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.

*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

PB9181 is reactive to FHIT in Human, Rat

Applications

PB9181 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

17 kDa

Calculated molecular weight

16858 MW

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.

View more details

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, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

Click image to see more details

Figure 1. 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

Figure 2. 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

Figure 4. 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

Figure 3. 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.

Gene/Protein Information For FHIT (Source: Uniprot.org, NCBI)

Alternative Names

AP3A hydrolase; AP3AaseFRA3Bbis(5'-adenosyl)-triphosphatase; Diadenosine 5'-5'''-P1; dinucleosidetriphosphatase; EC 3.6.1.29; fragile histidine triad gene; Fragile histidine triad protein; P3-triphosphate hydrolase; tumor suppressor protein FHIT AP3Aase, FRA3B fragile histidine triad diadenosine triphosphatase bis(5-adenosyl)-triphosphatase|AP3A hydrolase|diadenosine 5,5-P1,P3-triphosphate hydrolase|dinucleosidetriphosphatase

For more info on FHIT, check out the FHIT 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 FHIT: 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].

Take me to the FHIT Infographic

Here are featured tools and databases that you might find useful.

• Boster's Pathways Library
• Protein Databases
• Bioscience Research Protocol Resources
• Data Processing & Analysis Software
• Photo Editing Software
• Scientific Literature Resources
• Research Paper Management Tools
• Molecular Biology Software
• Primer Design Tools
• Bioinformatics Tools
• Phylogenetic Tree Analysis