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Product Info Summary
| SKU: | A11989-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-DPH5 Antibody Picoband®
SKU/Catalog Number
A11989-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-DPH5 Antibody Picoband® catalog # A11989-1. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human. 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
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.
Cite This Product
Anti-DPH5 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A11989-1)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human DPH5 recombinant protein (Position: D11-H250).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A11989-1 is reactive to DPH5 in Human
Observed Molecular Weight
37 kDa
Calculated molecular weight
31.7 kDa
Background of DPH5
Diphthine synthase is an enzyme that in humans is encoded by the DPH5 gene. This gene encodes a component of the diphthamide synthesis pathway. Diphthamide is a post-translationally modified histidine residue found only on translation elongation factor 2. It is conserved from archaebacteria to humans, and is targeted by diphtheria toxin and Pseudomonas exotoxin A to halt cellular protein synthesis. The yeast and Chinese hamster homologs of this protein catalyze the trimethylation of the histidine residue on elongation factor 2, resulting in a diphthine moiety that is subsequently amidated to yield diphthamide. Multiple transcript variants encoding different isoforms have been found for this gene.
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
A11989-1 is guaranteed for ELISA, Flow Cytometry, IF, 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, 0.25-0.5 μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human 293T whole cell, human HepG2 whole cell, human K562 whole cell, human Daudi whole cell
ICC/IF: U20S cell
FCM: A549 cell
Validation Images & Assay Conditions
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Western blot analysis of DPH5 using anti-DPH5 antibody (A11989-1).
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 293T whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human Daudi whole cell 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-DPH5 antigen affinity purified polyclonal antibody (Catalog # A11989-1) 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 DPH5 at approximately 37 kDa. The expected band size for DPH5 is at 32 kDa.
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IF analysis of DPH5 using anti-DPH5 antibody (A11989-1).
DPH5 was detected in an 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 5 μg/mL rabbit anti-DPH5 Antibody (A11989-1) 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.
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Flow Cytometry analysis of A549 cells using anti-DPH5 antibody (A11989-1).
Overlay histogram showing A549 cells stained with A11989-1 (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-DPH5 Antibody (A11989-1, 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-DPH5 Antibody Picoband® (A11989-1)
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