HAF017). A specific band was detected for EGFR at approximately 175 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox"> HAF017). A specific band was detected for EGFR at approximately 175 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and MDA-MB-231 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for EGFR at approximately 175 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins." class="big_lightbox"> AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins." class="big_thumb" />

NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox"> NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_lightbox"> VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to plasma membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_thumb" />

AF231) at 3ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (Catalog # VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to keratinocytes. " class="big_lightbox"> AF231) at 3ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-goat IgG VisUCyte HRP Polymer Antibody (Catalog # VC004) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to keratinocytes. " class="big_thumb" />

1095-ER) was serially diluted and captured by Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)" class="big_lightbox"> 1095-ER) was serially diluted and captured by Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)" class="big_thumb" />

1095-ER) was serially diluted and captured by Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)" class="big_lightbox"> 1095-ER) was serially diluted and captured by Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF231) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)" class="big_thumb" />

HAF008). A specific band was detected for Phospho-EGFR (Y1173) at approximately 185 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox"> HAF008). A specific band was detected for Phospho-EGFR (Y1173) at approximately 185 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 µM pervanadate (PV) for 10 minutes. PVDF membrane was probed with 0.2 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody, followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-EGFR (Y1173) at approximately 185 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox"> NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

CTS005) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections." class="big_lightbox"> CTS005) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections." class="big_thumb" />

VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membrane in cancer cells. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_lightbox"> VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membrane in cancer cells. Staining was performed using our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents." class="big_thumb" />

236-EG; filled histogram) then stained with Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with para-formaldehyde and permeabilized with saponin." class="big_lightbox"> 236-EG; filled histogram) then stained with Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with para-formaldehyde and permeabilized with saponin." class="big_thumb" />

236-EG) for 5 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-EGFR (Y1173) at approximately 265 kDa (as indicated) using 2 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system." class="big_lightbox"> 236-EG) for 5 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-EGFR (Y1173) at approximately 265 kDa (as indicated) using 2 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y1173) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1095). This experiment was conducted under reducing conditions and using the 66-440 kDa separation system." class="big_thumb" />

HAF019). A specific band was detected for EGFR at approximately 170 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox"> HAF019). A specific band was detected for EGFR at approximately 170 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and MDA-MB-231 human breast cancer cell line. PVDF membrane was probed with 0.25 µg/mL of Goat Anti-Mouse EGFR Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1280) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for EGFR at approximately 170 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to developing muscle. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections." class="big_lightbox"> CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to developing muscle. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections." class="big_thumb" />

HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." class="big_lightbox"> HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." class="big_thumb" />

236-EG) for 5 minutes. PVDF membrane was probed with 1 µg/mL of Human Phospho-EGFR (Y1068) Monoclonal Antibody (Catalog # MAB3570), followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-EGFR (Y1068) at approximately 190 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox"> 236-EG) for 5 minutes. PVDF membrane was probed with 1 µg/mL of Human Phospho-EGFR (Y1068) Monoclonal Antibody (Catalog # MAB3570), followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-EGFR (Y1068) at approximately 190 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human EGF (Catalog # 236-EG) for 5 minutes. PVDF membrane was probed with 1 µg/mL of Human Phospho-EGFR (Y1068) Monoclonal Antibody (Catalog # MAB3570), followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-EGFR (Y1068) at approximately 190 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

236-EG, filled histogram) then stained with Human Phospho--EGFR (Y1068) Monoclonal Antibody (Catalog # MAB3570), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0102B). Mouse IgG_2A(Catalog # MAB003, data not shown) was used as an isotype control. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin." class="big_lightbox"> 236-EG, filled histogram) then stained with Human Phospho--EGFR (Y1068) Monoclonal Antibody (Catalog # MAB3570), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0102B). Mouse IgG_2A(Catalog # MAB003, data not shown) was used as an isotype control. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin." class="big_thumb" />

236-EG) for 5 minutes to induce tyrosine phosphorylation of EGF R. Lysates were serially diluted and analyzed by (A) IP-Western blot and (B) this DuoSet IC ELISA. IPs were done using an anti-EGF R monoclonal antibody and anti-mouse IgG agarose. Immunoblots were incubated with an HRP-conjugated anti-phospho-tyrosine monoclonal antibody (R&D Systems, Catalog #HAM1676) to detect phosphor-EGF R. Bands were visualized by chemiluminescent detection. Human Phospho-EGF R can be detected in this DuoSet IC ELISA by using approximately 2-4 times less lysate than is needed for a conventional IP-Western blot." class="big_lightbox"> 236-EG) for 5 minutes to induce tyrosine phosphorylation of EGF R. Lysates were serially diluted and analyzed by (A) IP-Western blot and (B) this DuoSet IC ELISA. IPs were done using an anti-EGF R monoclonal antibody and anti-mouse IgG agarose. Immunoblots were incubated with an HRP-conjugated anti-phospho-tyrosine monoclonal antibody (R&D Systems, Catalog #HAM1676) to detect phosphor-EGF R. Bands were visualized by chemiluminescent detection. Human Phospho-EGF R can be detected in this DuoSet IC ELISA by using approximately 2-4 times less lysate than is needed for a conventional IP-Western blot." alt="The human epidermoid carcinoma cell line, A431, was treated with 25 ng/mL recombinant human EGF (R&D Systems Catalog #236-EG) for 5 minutes to induce tyrosine phosphorylation of EGF R. Lysates were serially diluted and analyzed by (A) IP-Western blot and (B) this DuoSet IC ELISA. IPs were done using an anti-EGF R monoclonal antibody and anti-mouse IgG agarose. Immunoblots were incubated with an HRP-conjugated anti-phospho-tyrosine monoclonal antibody (R&D Systems, Catalog #HAM1676) to detect phosphor-EGF R. Bands were visualized by chemiluminescent detection. Human Phospho-EGF R can be detected in this DuoSet IC ELISA by using approximately 2-4 times less lysate than is needed for a conventional IP-Western blot." class="big_thumb" />

AF231)." class="big_lightbox"> AF231)." class="big_thumb" />

1095-ER), human ErbB2/Fc Chimera (R&D Systems, Catalog #1129-ER), human ErbB3/Fc Chimera (R&D Systems, Catalog #348-RB), or human ErbB4/Fc Chimera (R&D Systems, Catalog #1131-ER) were added to 25 μg of lysate and analyzed using this DuoSet IC ELISA. Competition was observed only with recombinant human EGF R." class="big_lightbox"> 1095-ER), human ErbB2/Fc Chimera (R&D Systems, Catalog #1129-ER), human ErbB3/Fc Chimera (R&D Systems, Catalog #348-RB), or human ErbB4/Fc Chimera (R&D Systems, Catalog #1131-ER) were added to 25 μg of lysate and analyzed using this DuoSet IC ELISA. Competition was observed only with recombinant human EGF R." class="big_thumb" />

344-ER) binds recombinant human EGF (236-EG) in a functional ELISA. The estimated K_d for this interaction is < 8 nM." class="big_lightbox"> 344-ER) binds recombinant human EGF (236-EG) in a functional ELISA. The estimated K_d for this interaction is < 8 nM." class="big_thumb" />

AF231) Immunoblots were incubated with a biotinylated anti-EGF R polyclonal antibody (Catalog # BAF231) to detect total EGF R. Blots were incubated with Streptavidin-HRP (Catalog # DY998) followed by chemiluminescent detection. Human EGF R can be detected by the Human Total-EGF R DuoSet IC ELISA by using approximately 200 to 500 times less lysate than is needed for a conventional IP-Western blot. " class="big_lightbox"> AF231) Immunoblots were incubated with a biotinylated anti-EGF R polyclonal antibody (Catalog # BAF231) to detect total EGF R. Blots were incubated with Streptavidin-HRP (Catalog # DY998) followed by chemiluminescent detection. Human EGF R can be detected by the Human Total-EGF R DuoSet IC ELISA by using approximately 200 to 500 times less lysate than is needed for a conventional IP-Western blot. " class="big_thumb" />

236-EG) for 5 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y845) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3394), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-EGFR (Y845) at approximately 175 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox"> 236-EG) for 5 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y845) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3394), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-EGFR (Y845) at approximately 175 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human EGF (Catalog # 236-EG) for 5 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-EGFR (Y845) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3394), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-EGFR (Y845) at approximately 175 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

236-EG; filled histogram) then stained with Rabbit Anti-Human Phospho-EGFR (Y845) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3394), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). Rabbit IgG (Catalog # AB-105-C, data not shown) was used as a control antibody. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin." class="big_lightbox"> 236-EG; filled histogram) then stained with Rabbit Anti-Human Phospho-EGFR (Y845) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3394), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). Rabbit IgG (Catalog # AB-105-C, data not shown) was used as a control antibody. To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin." class="big_thumb" />

NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox"> NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

236-EG) for 5 minutes. PVDF membrane was probed with 1 µg/mL of Rat Anti-Human Phospho-EGFR (Y1086) Monoclonal Antibody (Catalog # MAB89671) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Phospho-EGFR (Y1086) at approximately 180 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_lightbox"> 236-EG) for 5 minutes. PVDF membrane was probed with 1 µg/mL of Rat Anti-Human Phospho-EGFR (Y1086) Monoclonal Antibody (Catalog # MAB89671) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Phospho-EGFR (Y1086) at approximately 180 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." alt="Western blot shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 ng/mL Recombinant Human EGF (Catalog # 236-EG) for 5 minutes. PVDF membrane was probed with 1 µg/mL of Rat Anti-Human Phospho-EGFR (Y1086) Monoclonal Antibody (Catalog # MAB89671) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Phospho-EGFR (Y1086) at approximately 180 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." class="big_thumb" />

236-EG) using Rat Anti-Human EGFR Monoclonal Antibody (Catalog # MAB89671) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cell membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_lightbox"> 236-EG) using Rat Anti-Human EGFR Monoclonal Antibody (Catalog # MAB89671) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cell membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." class="big_thumb" />

236-EG) for 5 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-EGFR (Y1086) at approximately 197 kDa (as indicated) using 10 µg/mL of Rat Anti-Human Phospho-EGFR (Y1086) Monoclonal Antibody (Catalog # MAB89671) followed by 1:50 dilution of HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." class="big_lightbox"> 236-EG) for 5 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-EGFR (Y1086) at approximately 197 kDa (as indicated) using 10 µg/mL of Rat Anti-Human Phospho-EGFR (Y1086) Monoclonal Antibody (Catalog # MAB89671) followed by 1:50 dilution of HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system." class="big_thumb" />

F0135). View our protocol for Staining Membrane-associated Proteins." class="big_lightbox"> F0135). View our protocol for Staining Membrane-associated Proteins." alt="A431 human epithelial carcinoma cell line was stained with Human Anti-Human EGF R/ErbB1 (Research Grade Cetuximab Biosimilar) Monoclonal Antibody (Catalog # MAB9577, filled histogram) or irrelevant antibody (open histogram) followed by APC-conjugated Anti-Human IgG Secondary Antibody (Catalog # F0135). View our protocol for Staining Membrane-associated Proteins." class="big_thumb" />