Anti-EGFR (phospho Y1045) antibody (ab277783)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit recombinant multiclonal [1HCLC] to EGFR (phospho Y1045)
- Suitable for: WB, ICC/IF
- Reacts with: Human
Overview
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Product name
Anti-EGFR (phospho Y1045) antibody [1HCLC]
See all EGFR primary antibodies -
Description
Rabbit recombinant multiclonal [1HCLC] to EGFR (phospho Y1045) -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide aa 1066-1076 (phospho Y1045).
Database link: P00533 -
Positive control
- WB: A-431 untreated and treated with EGF (10 ng/mL for 5 min). ICC/IF: A-431 cells.
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General notes
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains.
Recombinant multiclonal antibodies offer the sensitivity of polyclonal antibodies by recognising multiple epitopes, along with consistency of a recombinant antibody.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.09% Sodium azide
Constituent: 99.91% PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Recombinant Multiclonal -
Clone number
1HCLC -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-EGFR antibody (ab277783)All lanes : Anti-EGFR (phospho Y1045) antibody (ab277783) at 2 µg/ml
Lane 1 : A431 (human epidermoid carcinoma cell line) whole cell extract
Lane 2 : A431 (human epidermoid carcinoma cell line) whole cell extract (treated with EGF 10 ng / mL for 5 minutes)
Predicted band size: 134 kDa
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Immunocytochemistry - Anti-EGFR antibody (ab277783)For immunofluorescence analysis A431 cells were fixed and permeabilized for detection of EGFR pY1045 using Anti-EGFR pY1045 Recombinant Rabbit Multiclonal Antibody (ab277783, 2 μ/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (1/2000). Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI, and Alexa Fluor® 555 Rhodamine Phalloidin (1/300) was used for cytoskeletal F-actin (red) staining. Detection and localization of EGFR pY1045 (green) in the membrane/cytoplasm can be clearly observed in cells treated with EGF (100 ng/mL, 15 min) as compared to untreated cells. Antibody specificity was demonstrated by competition with the EGFR pY1045 peptide, which results in loss of signal. No competition was observed with the non-phospho peptide. The images were captured at 60X magnification.
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Immunocytochemistry - Anti-EGFR antibody (ab277783)Immunofluorescence analysis of Phospho-EGFR (Tyr1045) was performed using 70% confluent log phase A-431 cells treated with 200 ng/mL of EGF for 10 minutes. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-EGFR (Tyr1045) Recombinant Rabbit Multiclonal Antibody (ab277783) at 2 μ/mL in 0.1% BSA and incubated overnight at +4°C and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate at a dilution of 1/2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI. F-actin (Panel c: red) was stained with Rhodamine Phalloidin (1/300). Panel d represents the merged image showing membrane localization. Panel e represents cells treated with antagonist, Afatinib (1uM for 6hrs) followed by EGF (200 ng/mL for 10 minutes), showing no Phospho-EGFR staining. Panel f shows untreated cells with no signal. Panel g represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
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Anti-EGFR antibody (ab277783)