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Product Info Summary
| SKU: | PB9016 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Rat |
| Host: | Rabbit |
| Application: | IF, ICC, WB |
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Product info
Product Name
Anti-EGFR Antibody Picoband®
SKU/Catalog Number
PB9016
PB0039 is an alternative SKU for this antibody, used in previous lots.
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-EGFR Antibody Picoband® catalog # PB9016. Tested in IF, ICC, WB applications. This antibody reacts with Human, 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-EGFR Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9016)
Host
Rabbit
Contents
Each vial contains antibody formulated with stabilizing components, 0.9 mg NaCl, 0.2 mg Na2HPO4, and 0.05 mg NaN3.
*This antibody is supplied in a stabilized formulation.
Compatibility with conjugation reactions depends on the chemistry of the conjugation method used.
For conjugation methods that are not compatible with the stabilizing components present in this formulation, a carrier-free antibody format is required.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human EGFR recombinant protein (Position: L25-K346). Human EGFR shares 89% amino acid (aa) sequence identity with mouse EGFR.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9016 is reactive to EGFR in Human, Rat
Observed Molecular Weight
200 kDa
Calculated molecular weight
134.3 kDa
Background of EGFR
The epidermal growth factor receptor (EGFR; ErbB-1; HER1 in humans) is the cell-surface receptor for members of the epidermal growth factor family (EGF-family) of extracellular protein ligands. It is a member of the ErbB family of receptors, a subfamily of four closely related receptor tyrosine kinases: EGFR (ErbB-1), HER2/c-neu (ErbB-2), Her 3 (ErbB-3) and Her 4 (ErbB-4). EGFR exists on the cell surface and is activated by binding of its specific ligands, including epidermal growth factor and transforming growth factor alpha (TGFalpha). EGFR and its ligands are cell signaling molecules involved in diverse cellular functions, including cell proliferation, differentiation, motility, and survival, and in tissue development. Mutations that lead to EGFR overexpression (known as upregulation) or overactivity have been associated with a number of cancers, including lung cancer and glioblastoma multiforme. In this latter case a more or less specific mutation of EGFR, called EGFRvIII is often observed.
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
PB9016 is guaranteed for 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, 0.1-0.5μg/ml, Human, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Positive Control
WB: HELA Whole Cell, A549 Whole Cell, A431 Whole Cell
ICC/IF: A431 cell
Validation Images & Assay Conditions
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Western blot analysis of EGFR using anti-EGFR antibody (PB9016).
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: HELA Whole Cell Lysate,
Lane 2: A549 Whole Cell Lysate,
Lane 3: A431 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-EGFR antigen affinity purified polyclonal antibody (Catalog # PB9016) 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 EGFR at approximately 200KD. The expected band size for EGFR is at 134KD.
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IF analysis of EGFR using anti-EGFR antibody (PB9016).
EGFR was detected in immunocytochemical section of A431 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-EGFR Antibody (PB9016) 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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Western blot analysis of EGFR using anti-EGFR antibody (PB9016).
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.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% milk in PBS/0.05% Tween-20 (5% milk/PBS/Tw) for 1.5 hour at RT. The membrane was incubated with rabbit anti-EGFR antibody (PB9016) at 1 ug/mL in 5% milk/PBS/0.05% Tween 20 overnight at 4℃ , then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit antibody conjugated with HRP at 1:5,000 in 5% milk/PBS/Tw at 4℃ for 12 hours. The signal is developed using an SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific). A specific band was detected for EGFR at approximately 134 kDa. The expected band size for EGFR is at 134 kDa.
Specific Publications For Anti-EGFR Antibody Picoband® (PB9016)
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Customer Reviews
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1 Reviews For Anti-EGFR Antibody Picoband®
I would highly recommend this Boster antibody...--Elena Deryugina, CELL AND MOLECULAR BIOLOGY, THE SCRIPPS RESEARCH INSTITUTE, ASSISTANT PROFESSOR
Excellent
Source: Biocompare.com
| SKU | PB9016 |
|---|---|
| Applications | Western Blot |
| Sample | Recombinant N-Terminal domain of human EGFR (aa 1-621) tested at 100 ng/lane |
| Detection | SuperSignal West Pico Chemiluminescent Substrate |
"Rabbit antibody PB9016 from Boster was probed for western blot detection of the recombinant N-terminal domain of human EGFR (100 ng/lane). The antibody nicely detected the protein band with the expected mol.wt. of recombinant protein. The antibody appears to be specific since non-specific bands were not detected in parallel lanes with cell lysates lacking EGFR. I would highly recommend this Boster antibody for detection of EGFR with the intact N-terminal domain."
Elena Deryugina
Verified customer
Submitted 2016-08-24
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