Simple Western



Ku80/XRCC5 Antibody (5D5.8) - NB100-103 from Novus Biologicals

Host Mouse
Clonality Monoclonal
Clone 5D5.8
Western Blot: Ku80 Antibody (5D5.8) [NB100-103] - WB analysis of Ku80 in MCF7 cell lysate using NB100-103.

ABCG2/CD338 Antibody - NBP1-59749 from Novus Biologicals

Host Rabbit
Clonality Polyclonal
Western Blot: ABCG2 Antibody [NBP1-59749] - Titration: 2.5 ug/ml Positive Control: HepG2 cell lysate.

FAK [p tyr397] Antibody - AF4528 from Novus Biologicals

Host Rabbit
Clonality Polyclonal
Western Blot: FAK Antibody [AF4528] - Lysates of HUVEC human umbilical vein endothelial cell untreated (-) or treated (+) with 100 uM pervanadate (PV) for 10 minutes. PVDF membrane was probed with 1 ug/mL of Human Phospho-FAK (Y397) Antigen Affinity-purified Polyclonal Antibody (R&D Systems catalog # AF4528), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (R&D Systems catalog # HAF008). A specific band was detected for Phospho-FAK (Y397) at approximately 122 kDa (as indicated). This experiment was conducted under reducing conditions.

His Tag MAb (Clone AD1.1.10), Mouse IgG1 - MAB050-SP from R&D Systems

Host Mouse
Clonality Monoclonal
Clone AD1.1.10
Detection of His Tag by Simple WesternTM. Simple Western lane view shows lysates of CHO Chinese hamster ovary cell line transfected with His-tagged Human S1P4/EDG-6, loaded at 0.5 mg/mL. A specific band was detected for His-tagged S1P4/EDG-6 at approximately 75 kDa (as indicated) using 10 µg/mL of Mouse Anti-His Tag Monoclonal Antibody (Catalog # MAB050). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Human SOX17 Affinity Purified Polyclonal Ab, Goat IgG - AF1924-SP from R&D Systems

Host Goat
Clonality Polyclonal
Detection of Human SOX17 by Western Blot. Western blot shows lysates of BG01V human embryonic stem cells untreated (-) or endoderm differentiated (+). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human SOX17 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1924) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for SOX17 at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Human SOX2 Affinity Purified Polyclonal Ab, Goat IgG - AF2018-SP from R&D Systems

Host Goat
Clonality Polyclonal
Detection of Human SOX2 by Western Blot. Western blot shows lysates of BG01V human embryonic stem cells and NTera‑2 human testicular embryonic carcinoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human SOX2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2018) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for SOX2 at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Human/Mouse SOX2 MAb (Clone 245610), Mouse IgG2A - MAB2018-SP from R&D Systems

Host Mouse
Clonality Monoclonal
Clone 245610
E‑Cadherin and SOX2 in BG01V Human Stem Cells. E‑Cadherin and SOX2 were detected in BG01V human embryonic stem cells using 10 µg/mL Goat Anti-Human E‑Cadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF648) and 10 µg/mL Mouse Anti-Human/Mouse SOX2 Monoclonal Antibody (Catalog # MAB2018). Cells were incubated with primary antibodies for 3 hours at room temperature. Cells were stained for E‑Cadherin using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (green; Catalog # NL001) and for SOX2 using the NorthernLights 493-conjugated Anti-Mouse Secondary Antibody (red; Catalog # NL009). Cells were counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Neuron-specific beta-III Tubulin MAb (Clone TuJ-1), Mouse IgG2A - MAB1195-SP from R&D Systems

Host Mouse
Clonality Monoclonal
Clone TuJ-1
beta ‑III Tubulin in Differentiated Human Neural Progenitor Cells. beta ‑III Tubulin was detected in immersion fixed differentiated human neural progenitor cells using Neuron-specific beta ‑III Tubulin Monoclonal Antibody (clone TuJ-1) (Catalog # MAB1195) for 3 hours at room temperature. Cells were stained (green) and counterstained (red). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

His Tag HRP MAb (Clone AD1.1.10), Mouse IgG1 - MAB050H-SP from R&D Systems

Host Mouse
Clonality Monoclonal
Clone AD1.1.10
Detection of His Tag by Simple WesternTM. Simple Western lane view shows His-tagged Recombinant Human BAI1 (Catalog # 4969-BA), loaded at 5 µg/mL. A specific band was detected for His-tagged Recombinant Human BAI1 at approximately 153 kDa (as indicated) using 1:50 dilution of Mouse Anti-His Tag HRP-conjugated Monoclonal Antibody (Catalog # MAB050H). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Human/Mouse 4EBP1 Affinity Purified Polyclonal Ab, Goat IgG - AF3227-SP from R&D Systems

Host Goat
Clonality Polyclonal
Detection of Human/Mouse 4EBP1 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and C2C12 mouse myoblast cell line. PVDF membrane was probed with 0.5 µg/mL of Human/Mouse 4EBP1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3227) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). Specific bands were detected for 4EBP1 at approximately 18-20 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Phospho-EGF R/ErbB1 (Y1173) by Western Blot. 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-EGF R/ErbB1 (Y1173) Antigen Affinity-purified Polyclonal Antibody, followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-EGF R/ErbB1 (Y1173) at approximately 185 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Human PDX-1/IPF1 Affinity Purified Polyclonal Ab, Goat IgG - AF2419-SP from R&D Systems

Host Goat
Clonality Polyclonal
Detection of Human PDX‑1/IPF1 by Western Blot. Western blot shows lysates of beta TC‑6 mouse beta cell insulinoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human PDX‑1/IPF1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2419) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for PDX‑1/IPF1 at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Detection of Human HIF‑1 alpha by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line untreated (-) or treated (+) with 150 µM CoCl2 for 16 hours. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat HIF‑1 alpha Antigen-affinity Purified Polyclonal Antibody, followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for HIF‑1 alpha at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Human/Mouse HSP27 Affinity Purified Polyclonal Ab, Rabbit IgG - AF1580-SP from R&D Systems

Host Rabbit
Clonality Polyclonal
Detection of Human/Mouse HSP27 by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line, HeLa human cervical epithelial carcinoma cell line, and C2C12 mouse myoblast cell line. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse HSP27 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1580) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for HSP27 at approximately 27 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Human/Mouse/Rat HIF-1 alpha MAb (Clone 241809), Mouse IgG1 - MAB1536-SP from R&D Systems

Host Mouse
Clonality Monoclonal
Clone 241809
Detection of Human, Mouse, and Rat HIF‑1 alpha by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line, Balb‑3T3 mouse embryonic fibroblast cell line, and PC‑12 rat adrenal pheochromocytoma cell line untreated (-) or treated (+) with 150 µM CoCl2 for 8 hours. PVDF membrane was probed with 1 µg/mL of Human/Mouse/Rat HIF‑1 alpha Monoclonal Antibody, followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for HIF‑1 alpha at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Human SPARC Affinity Purified Polyclonal Ab, Goat IgG - AF941-SP from R&D Systems

Host Goat
Clonality Polyclonal
Detection of Human SPARC by Western Blot. Western blot shows lysates of MG‑63 human osteosarcoma cell line. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human SPARC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF941) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for SPARC at approximately 43 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Lipocalin‑2/NGAL by Western Blot. Western blot shows lysates of Capan‑1 human pancreatic adenocarcinoma cell line and human pancreas tissue. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Human Lipocalin‑2/NGAL Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1757) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Lipocalin‑2/NGAL at approximately 22 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Human Aurora A Affinity Purified Polyclonal Ab, Goat IgG - AF3295-SP from R&D Systems

Host Goat
Clonality Polyclonal
Detection of Human Aurora A by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line. PVDF membrane was probed with 0.25 µg/mL of Human Aurora A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3295) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Aurora A at approximately 54 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 5.

Human Cadherin-11 MAb (Clone 283416), Mouse IgG2B - MAB1790-SP from R&D Systems

Host Mouse
Clonality Monoclonal
Clone 283416
Detection of Human Cadherin‑11 by Western Blot.Western blot shows lysates of PC‑3 human prostate cancer cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Cadherin‑11 Monoclonal Antibody (Catalog # MAB1790) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Cadherin‑11 at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human/Mouse Myeloperoxidase/MPO by Western Blot. Western blot shows lysates of HL‑60 human acute promyelocytic leukemia cell line, human neutrophil cells, and mouse spleen tissue. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse Myeloperoxidase/MPO Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3667) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Myeloperoxidase/MPO at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.

Human Galectin-1 Affinity Purified Polyclonal Ab, Goat IgG - AF1152-SP from R&D Systems

Host Goat
Clonality Polyclonal
Detection of Human Galectin‑1 by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and A549 human lung carcinoma cell line. PVDF membrane was probed with 0.1 µg/mL of Goat Anti-Human Galectin‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1152) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Galectin‑1 at approximately 14 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Human Fibronectin MAb (Clone P1H11), Mouse IgG1 - MAB1918-SP from R&D Systems

Host Mouse
Clonality Monoclonal
Clone P1H11
Fibronectin in Human Breast Cancer Tissue. Fibronectin was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Mouse Anti-Human Fibronectin Monoclonal Antibody (Catalog # MAB1918) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to connective tissue and fibroblasts. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Human/Mouse alpha-Fetoprotein MAb (Clone 189502), Mouse IgG1 - MAB1368-SP from R&D Systems

Host Mouse
Clonality Monoclonal
Clone 189502
Detection of Human alpha ‑Fetoprotein/AFP by Simple WesternTM. Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for alpha ‑Fetoprotein/AFP at approximately 70 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human/Mouse alpha ‑Fetoprotein/AFP Monoclonal Antibody (Catalog # MAB1368). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Human and Mouse Phospho-p70 S6 Kinase (T421/S424) by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line untreated (-) or treated (+) with 100 ng/mL of Recombinant Human IGF‑1 (Catalog # 291-G1) for 20 minutes and NIH‑3T3 mouse embryonic fibroblast cell line untreated or treated with 100 ng/mL of Human PDGF (Catalog # 120-HD) for 20 minutes. PVDF membrane was probed with 0.25 µg/mL of Human/Mouse/Rat Phospho-p70 S6 Kinase (T421/S424) Antigen Affinity-purified Polyclonal Antibody, followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for Phospho-p70 S6 Kinase (T421/S424) at approximately 70 and 85 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Human Myeloperoxidase Affinity Purified Polyclonal Ab, Goat IgG - AF3174-SP from R&D Systems

Host Goat
Clonality Polyclonal
Detection of Human Myeloperoxidase/MPO by Western Blot. Western blot shows lysates of HL‑60 human acute promyelocytic leukemia cell line and human neutrophil. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human Myeloperoxidase/MPO Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3174) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Myeloperoxidase/MPO at approximately 60-65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
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