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Horseradish peroxidase conjugated goat anti mouse and goat anti rabbit secondary antibodies

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Horseradish-peroxidase conjugated goat anti-mouse and goat anti-rabbit secondary antibodies are laboratory reagents used in immunoassays and immunohistochemistry applications. They are designed to detect and amplify the signal from primary antibodies that bind to target molecules in samples.

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Lab products found in correlation

Reducing sample buffer, by Carl Roth (5 mentions) β actin, by Abcam (5 mentions)

6 protocols using horseradish peroxidase conjugated goat anti mouse and goat anti rabbit secondary antibodies

1

CLDN6 Protein Expression in Cancers

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Example 2

For Western blot analysis 20 μg of total protein extracted from cells lyzed with Laemmli-lysis buffer was used. Extracts were diluted in reducing sample buffer (Roth), subjected to SDS-PAGE and subsequently electrotransferred onto PVDF membrane (Pall). Immunostaining was performed with polyclonal antibodies reactive to CLDN6 (ARP) and beta-Actin (Abcam) followed by detection of primary antibodies with horseradish-peroxidase conjugated goat anti-mouse and goat anti-rabbit secondary antibodies (Dako).

Tissue lysates from up to five individuals were tested for each normal tissue type. No CLDN6 protein expression was detected in any of the normal tissues analyzed. In contrast to normal tissues, high expression of CLDN6 protein was detected in samples from ovarian cancer and lung cancer. CLDN6 expression was detected in NIH-OVCAR3 (ovarian cancer), MKN7 (gastric cancer), AGS (gastric cancer), CPC-N(SCLC), HCT-116 (colon cancer), FU97 (gastric cancer), NEC8 (testicular embryonal carcinoma), JAR (placental choriocarcinoma), JEG3 (placental choriocarcinoma), BEWO (placental choriocarcinoma), and PA-1 (ovarian teratocarcinoma).

US10919974B2. Antibodies for treatment of cancer expressing claudin 6 (2021-02-16). Ganymed Pharmaceuticals AG [DE], Johannes Gutenberg-Universitat Mainz [DE]. Inventors: Ugur Sahin [DE], Ozlem Tureci [DE], Michael Koslowski [DE], Korden Walter [DE], Stefan Woll [DE], Maria Kreuzberg [DE], Bernd Hubner [DE], Michael Erdeljan [DE], Michael Weichel [DE].
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2

Western Blot Analysis of CLDN6 Expression

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Example 2

For Western blot analysis 20 μg of total protein extracted from cells lyzed with Laemmli-lysis buffer was used. Extracts were diluted in reducing sample buffer (Roth), subjected to SDS-PAGE and subsequently electrotransferred onto PVDF membrane (Pall). Immunostaining was performed with polyclonal antibodies reactive to CLDN6 (ARP) and beta-Actin (Abcam) followed by detection of primary antibodies with horseradish-peroxidase conjugated goat anti-mouse and goat anti-rabbit secondary antibodies (Dako).

Tissue lysates from up to five individuals were tested for each normal tissue type. No CLDN6 protein expression was detected in any of the normal tissues analyzed. In contrast to normal tissues, high expression of CLDN6 protein was detected in samples from ovarian cancer and lung cancer. CLDN6 expression was detected in NIH-OVCAR3 (ovarian cancer), MKN7 (gastric cancer), AGS (gastric cancer), CPC-N(SCLC), HCT-116 (colon cancer), FU97 (gastric cancer), NEC8 (testicular embryonal carcinoma), JAR (placental choriocarcinoma), JEG3 (placental choriocarcinoma), BEWO (placental choriocarcinoma), and PA-1 (ovarian teratocarcinoma).

US09932401B2. Antibodies specific for claudin 6 (CLDN6) (2018-04-03). Ganymed Pharmaceuticals AG [DE], Johannes Gutenberg-Universitat Mainz [DE]. Inventors: Ugur Sahin [DE], Özlem Türeci [DE], Michael Koslowski [DE], Korden Walter [DE], Stefan Woll [DE], Maria Kreuzberg [DE], Bernd Hubner [DE], Michael Erdeljan [DE].
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3

Protein Quantification and Western Blot Analysis

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Total cellular protein was quantified by Lowry et al. (1951) (link) method and 100 μg of protein from each sample was loaded onto 10% gel separated on SDS-PAGE and nitrocellulose membrane was used to transfer proteins using electro blotting apparatus (Cleaver Scientific Ltd, UK) according to previous method (Anand et al., 2012 (link)). The membranes were probed with β-actin (ab8229), JNK3 (55A8), phospho-SAPK/JNK (Thr183/Tyr185; 98F2), c-jun (ab32137), phospho c-jun (phospho S63; ab32385) at 1: 1000 dilutions whereas active caspase 3 antibody (ab90437) at 1:400 dilution and incubated at 37°C for 3 h after transfer. TBST was used to wash the membranes four times for 15 min and incubated at room temperature for 2 h in horseradish peroxidase conjugated goat anti-mouse and goat anti-rabbit secondary antibodies (DAKO, Denmark) at a dilution of 1:10,000. The membranes were developed using an enhanced chemiluminescence detection system (ProteoQwest®, Sigma) after washing. After developing the membranes they were exposed to x-ray film and band intensity captured and measured using NIH imageJ analysis software.
Bhat P.V., Pandareesh M.D., Khanum F, & Tamatam A. (2016). Cytotoxic Effects of Ochratoxin A in Neuro-2a Cells: Role of Oxidative Stress Evidenced by N-acetylcysteine. Frontiers in Microbiology, 7, 1142.
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4

Protein Expression Analysis of CLDN6

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Example 2

For Western blot analysis 20 μg of total protein extracted from cells lyzed with Laemmli-lysis buffer was used. Extracts were diluted in reducing sample buffer (Roth), subjected to SDS-PAGE and subsequently electrotransferred onto PVDF membrane (Pall). Immunostaining was performed with polyclonal antibodies reactive to CLDN6 (ARP) and beta-Actin (Abcam) followed by detection of primary antibodies with horseradish-peroxidase conjugated goat anti-mouse and goat anti-rabbit secondary antibodies (Dako).

Tissue lysates from up to five individuals were tested for each normal tissue type. No CLDN6 protein expression was detected in any of the normal tissues analyzed. In contrast to normal tissues, high expression of CLDN6 protein was detected in samples from ovarian cancer and lung cancer. CLDN6 expression was detected in NIH-OVCAR3 (ovarian cancer), MKN7 (gastric cancer), AGS (gastric cancer), CPC-N (SCLC), HCT-116 (colon cancer), FU97 (gastric cancer), NEC8 (testicular embryonal carcinoma), JAR (placental choriocarcinoma), JEG3 (placental choriocarcinoma), BEWO (placental choriocarcinoma), and PA-1 (ovarian teratocarcinoma).

US11859008B2. Antibodies for treatment of cancer expressing claudin 6 (2024-01-02). Ganymed Pharmaceuticals GmbH [DE], Johannes Gutenberg-Universitat Mainz [DE]. Inventors: Ugur Sahin [DE], Ozlem Tureci [DE], Michael Koslowski [DE], Korden Walter [DE], Stefan Woll [DE], Maria Kreuzberg [DE], Bernd Hubner [DE], Michael Erdeljan [DE], Michael Weichel [DE].
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5

Quantifying CLDN6 Protein Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 2

For Western blot analysis 20 μg of total protein extracted from cells lyzed with Laemmli-lysis buffer was used. Extracts were diluted in reducing sample buffer (Roth), subjected to SDS-PAGE and subsequently electrotransferred onto PVDF membrane (Pall). Immunostaining was performed with polyclonal antibodies reactive to CLDN6 (ARP) and beta-Actin (Abcam) followed by detection of primary antibodies with horseradish-peroxidase conjugated goat anti-mouse and goat anti-rabbit secondary antibodies (Dako).

Tissue lysates from up to five individuals were tested for each normal tissue type. No CLDN6 protein expression was detected in any of the normal tissues analyzed. In contrast to normal tissues, high expression of CLDN6 protein was detected in samples from ovarian cancer and lung cancer. CLDN6 expression was detected in NIH-OVCAR3 (ovarian cancer), MKN7 (gastric cancer), AGS (gastric cancer), CPC-N(SCLC), HCT-116 (colon cancer), FU97 (gastric cancer), NEC8 (testicular embryonal carcinoma), JAR (placental choriocarcinoma), JEG3 (placental choriocarcinoma), BEWO (placental choriocarcinoma), and PA-1 (ovarian teratocarcinoma).

US11858988B2. Antibodies specific for claudin 6 (CLDN6) (2024-01-02). Ganymed Pharmaceuticals GmbH [DE], Johannes Gutenberg-Universitat Mainz [DE]. Inventors: Ugur Sahin [DE], Özlem Türeci [DE], Michael Koslowski [DE], Korden Walter [DE], Stefan Woll [DE], Maria Kreuzberg [DE], Bernd Hubner [DE], Michael Erdeljan [DE].
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6

Western Blot Analysis of CLDN6 Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Example 2

For Western blot analysis 20 μg of total protein extracted from cells lyzed with Laemmli-lysis buffer was used. Extracts were diluted in reducing sample buffer (Roth), subjected to SDS-PAGE and subsequently electrotransferred onto PVDF membrane (Pall). Immunostaining was performed with polyclonal antibodies reactive to CLDN6 (ARP) and beta-Actin (Abcam) followed by detection of primary antibodies with horseradish-peroxidase conjugated goat anti-mouse and goat anti-rabbit secondary antibodies (Dako).

Tissue lysates from up to five individuals were tested for each normal tissue type. No CLDN6 protein expression was detected in any of the normal tissues analyzed. In contrast to normal tissues, high expression of CLDN6 protein was detected in samples from ovarian cancer and lung cancer. CLDN6 expression was detected in NIH-OVCAR3 (ovarian cancer), MKN7 (gastric cancer), AGS (gastric cancer), CPC-N(SCLC), HCT-116 (colon cancer), FU97 (gastric cancer), NEC8 (testicular embryonal carcinoma), JAR (placental choriocarcinoma), JEG3 (placental choriocarcinoma), BEWO (placental choriocarcinoma), and PA-1 (ovarian teratocarcinoma).

US10745477B2. Antibodies specific for claudin 6 (CLDN6) (2020-08-18). Ganymed Pharmaceuticals AG [DE], Johannes Gutenberg-Universitat Mainz [DE]. Inventors: Ugur Sahin [DE], Özlem Türeci [DE], Michael Koslowski [DE], Korden Walter [DE], Stefan Woll [DE], Maria Kreuzberg [DE], Bernd Hubner [DE], Michael Erdeljan [DE].
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