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Product Info Summary
| SKU: | A03676-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Monkey, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-ORC5L/ORC5 Antibody Picoband®
SKU/Catalog Number
A03676-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-ORC5L/ORC5 Antibody Picoband® catalog # A03676-1. Tested in WB applications. This antibody reacts with Human, Monkey, 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-ORC5L/ORC5 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03676-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence in the middle region of human ORC5, identical to the related mouse sequence.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A03676-1 is reactive to ORC5 in Human, Monkey, Mouse, Rat
Observed Molecular Weight
50 kDa
Calculated molecular weight
50.3 kDa
Background of ORC5
Origin recognition complex subunit 5 is a protein that in humans is encoded by the ORC5 (ORC5L) gene. The origin recognition complex (ORC) is a highly conserved six subunit protein complex essential for the initiation of the DNA replication in eukaryotic cells. Studies in yeast demonstrated that ORC binds specifically to origins of replication and serves as a platform for the assembly of additional initiation factors such as Cdc6 and Mcm proteins. The protein encoded by this gene is a subunit of the ORC complex. Alternatively spliced transcript variants encoding distinct isoforms have been described.
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
A03676-1 is guaranteed for 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, 0.1-0.25μg/ml, Human, Mouse, Rat, Monkey
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: monkey COS-7 whole cell, human placenta tissue, human K562 whole cell, human MCF-7 whole cell, rat testis tissue, rat kidney tissue, mouse ovary tissue, mouse testis tissue
ICC/IF: MCF-7 cell
FCM: U87 cell
Validation Images & Assay Conditions
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Western blot analysis of ORC5L/ORC5 using anti-ORC5L/ORC5 antibody (A03676-1).
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: monkey COS-7 whole cell lysates,
Lane 2: human placenta tissue lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human MCF-7 whole cell lysates,
Lane 5: rat testis tissue lysates,
Lane 6: rat kidney tissue lysates,
Lane 7: mouse ovary tissue lysates,
Lane 8: mouse testis 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-ORC5L/ORC5 antigen affinity purified polyclonal antibody (Catalog # A03676-1) at 0.25 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 ORC5L/ORC5 at approximately 50 kDa. The expected band size for ORC5L/ORC5 is at 50 kDa.
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IF analysis of ORC5L/ORC5 using anti-ORC5L/ORC5 antibody (A03676-1).
ORC5L/ORC5 was detected in an immunocytochemical section of MCF-7 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-ORC5L/ORC5 Antibody (A03676-1) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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 U87 cells using anti-ORC5L/ORC5 antibody (A03676-1).
Overlay histogram showing U87 cells stained with A03676-1 (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-ORC5L/ORC5 Antibody (A03676-1, 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-ORC5L/ORC5 Antibody Picoband® (A03676-1)
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