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Product Info Summary
| SKU: | A00174-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, WB |
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Product info
Product Name
Anti-Oct4/POU5F1 Antibody Picoband®
SKU/Catalog Number
A00174-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Oct4/POU5F1 Antibody catalog # A00174-1. Tested in ELISA, Flow Cytometry, WB applications. This antibody reacts with Human, Mouse, 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-Oct4/POU5F1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00174-1)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Oct4/POU5F1 recombinant protein (Position: E26-A300).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00174-1 is reactive to POU5F1 in Human, Mouse, Rat
Observed Molecular Weight
45 kDa
Calculated molecular weight
38.6 kDa
Background of POU5F1
Oct-4 (octamer-binding transcription factor 4), also known as POU5F1 (POU domain, class 5, transcription factor 1), is a protein that in humans is encoded by the POU5F1 gene. It is mapped to 6p21.33. This gene encodes a transcription factor containing a POU homeodomain that plays a key role in embryonic development and stem cell pluripotency. Aberrant expression of this gene in adult tissues is associated with tumorigenesis. This gene can participate in a translocation with the Ewing's sarcoma gene on chromosome 21, which also leads to tumor formation. Alternative splicing, as well as usage of alternative AUG and non-AUG translation initiation codons, results in multiple isoforms. One of the AUG start codons is polymorphic in human populations. Related pseudogenes have been identified on chromosomes 1, 3, 8, 10, and 12.
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
A00174-1 is guaranteed for ELISA, Flow Cytometry, 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, Mouse, Rat
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human Hela whole cell lysates, rat brain tissue lysates, mouse brain tissue lysates
FCM: A431 cell, HepG2 cell
Validation Images & Assay Conditions
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Western blot analysis of POU5F1 using anti-POU5F1 antibody (A00174-1).
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 Hela whole cell lysates,
Lane 2: rat brain tissue lysates,
Lane 3: mouse brain 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-POU5F1 antigen affinity purified polyclonal antibody (Catalog # A00174-1) 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 POU5F1 at approximately 45 kDa. The expected band size for POU5F1 is at 39 kDa.
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Flow Cytometry analysis of A431 cells using anti-POU5F1 antibody (A00174-1).
Overlay histogram showing A431 cells stained with A00174-1 (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 rabbit anti-POU5F1 Antibody (A00174-1, 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.
Specific Publications For Anti-Oct4/POU5F1 Antibody Picoband® (A00174-1)
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1 Customer Q&As for Anti-Oct4/POU5F1 Antibody Picoband®
Question
We are currently using anti-Oct4/POU5F1 antibody A00174-1 for rat tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it likely that the antibody can work on goat tissues as well?
Verified Customer
Verified customer
Asked: 2017-06-05
Answer
The anti-Oct4/POU5F1 antibody (A00174-1) has not been tested for cross reactivity specifically with goat 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 goat you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2017-06-05
