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Product Info Summary
| SKU: | A00023-5 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-EGFR Antibody Picoband®
SKU/Catalog Number
A00023-5
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-EGFR Antibody Picoband® catalog # A00023-5. Tested in ELISA, Flow Cytometry, IF, IHC, 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
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 # A00023-5)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human EGFR recombinant protein (Position: D1006-A1210).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00023-5 is reactive to EGFR in Human
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
180 kDa
Calculated molecular weight
134277 MW
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
A00023-5 is guaranteed for ELISA, Flow Cytometry, IF, IHC, 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, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human A431 whole cell, human A549 whole cell, human U87 whole cell, human HELA whole cell
IHC: human lung cancer tissue
ICC/IF: A431 cell
FCM: A549 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of EGFR using anti-EGFR antibody (A00023-5).
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: human A431 whole cell lysates,
Lane 2: human A549 whole cell lysates,
Lane 3: human U87 whole cell lysates,
Lane 4: human HELA whole cell lysates.
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 # A00023-5) 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 EGFR at approximately 180KD. The expected band size for EGFR is at 134KD.
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Figure 2. IHC analysis of EGFR using anti-EGFR antibody (A00023-5).
EGFR was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-EGFR Antibody (A00023-5) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
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Figure 3. Flow Cytometry analysis of A549 cells using anti-EGFR antibody (A00023-5).
Overlay histogram showing A549 cells stained with A00023-5 (Blue line).The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-EGFR Antibody (A00023-5, 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.
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Figure 4. IF analysis of EGFR using anti-EGFR antibody (A00023-5).
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 5μg/mL rabbit anti-EGFR Antibody (A00023-5) 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.
Protein Target Info & Infographic
Gene/Protein Information For EGFR (Source: Uniprot.org, NCBI)
Gene Name
EGFR
Full Name
Epidermal growth factor receptor
Weight
134277 MW
Superfamily
protein kinase superfamily
Alternative Names
Epidermal growth factor receptor; Egfr EGFR ERBB, ERBB1, ERRP, HER1, NISBD2, PIG61, mENA epidermal growth factor receptor epidermal growth factor receptor|avian erythroblastic leukemia viral (v-erb-b) oncogene homolog|cell growth inhibiting protein 40|cell proliferation-inducing protein 61|epidermal growth factor receptor tyrosine kinase domain|erb-b2 receptor tyrosine kinase 1|proto-oncogene c-ErbB-1|receptor tyrosine-protein kinase erbB-1
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on EGFR, check out the EGFR Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for EGFR: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-EGFR Antibody Picoband® (A00023-5)
Hello CJ!
A00023-5 has been cited in 2 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Epidermal Growth Factor Receptor in Cultured Human Retinal Pigment Epithelial Cells
Inhibition of angiogenesis involves in anticancer activity of riccardin D, a macrocyclic bisbibenzyl, in human lung carcinoma
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