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Product Info Summary
| SKU: | A00446-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, WB |
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Product info
Product Name
Anti-Raf1 Antibody Picoband®
SKU/Catalog Number
A00446-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Raf1 Antibody Picoband® catalog # A00446-1. Tested in 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-Raf1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00446-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived human Raf1 recombinant protein (Position: Y364-Q451). Human Raf1 shares 98.9% amino acid (aa) sequence identity with both mouse and rat Raf1.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00446-1 is reactive to RAF1 in Human, Mouse, Rat
Observed Molecular Weight
73 kDa
Calculated molecular weight
73.1 kDa
Background of RAF1
RAF proto-oncogene serine/threonine-protein kinase, also known as proto-oncogene c-RAF or simply c-Raf or even Raf-1, is an enzyme that in humans is encoded by the RAF1 gene. This gene is the cellular homolog of viral raf gene (v-raf). The encoded protein is a MAP kinase kinasekinase (MAP3K), which functions downstream of the Ras family of membrane associated GTPases to which it binds directly. Once activated, the cellular RAF1 protein can phosphorylate to activate the dual specificity protein kinases MEK1 and MEK2, which in turn phosphorylate to activate the serine/threonine specific protein kinases, ERK1 and ERK2. Activated ERKs are pleiotropic effectors of cell physiology and play an important role in the control of gene expression involved in the cell division cycle, apoptosis, cell differentiation and cell migration. Mutations in this gene are associated with Noonan syndrome 5 and LEOPARD syndrome 2.
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
A00446-1 is guaranteed for 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
Positive Control
WB: rat kidney tissue, mouse kidney tissue, human MCF-7 whole cell, human 22RV1 whole cell, human Hela whole cell
FCM: HeLa cell
Validation Images & Assay Conditions
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Western blot analysis of Raf1 using anti-Raf1 antibody (A00446-1).
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 50ug of sample under reducing conditions.
Lane 1: rat kidney tissue lysate,
Lane 2: mouse kidney tissue lysate,
Lane 3: human MCF-7 whole cell lysate,
Lane 4: human 22RV1 whole cell lysate,
Lane 5: human Hela whole cell lysate.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-Raf1 antigen affinity purified polyclonal antibody (Catalog # A00446-1) 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 1:10000 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 Raf1 at approximately 73KD. The expected band size for Raf1 is at 73KD.
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Flow Cytometry analysis of HeLa cells using anti-Raf1 antibody (A00446-1).
Overlay histogram showing HeLa cells stained with PB10089 (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-Raf1 Antibody (A00446-1,1μg/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.
Specific Publications For Anti-Raf1 Antibody Picoband® (A00446-1)
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1 Customer Q&As for Anti-Raf1 Antibody Picoband®
Question
We are currently using anti-Raf1 antibody A00446-1 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 possible that the antibody can work on zebrafish tissues as well?
Verified Customer
Verified customer
Asked: 2020-04-27
Answer
The anti-Raf1 antibody (A00446-1) has not been tested for cross reactivity specifically with zebrafish 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 zebrafish you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2020-04-27
