Paraffin-embedded tissue specimens (4 μm thick) were sectioned, dewaxed, and rehydrated in gradient ethanol. Then, we add 3% H2O2 and soak for 10 min to remove endogenous peroxidase. The antigen retrieval was accomplished in 10 mM citrate at 95°C for 20 min. Slides were blocked in goat serum for 60 min and incubated with rabbit anti-CAMK1D antibodies (1:100; ab172618; Abcam) overnight at 4°C. After washes in phosphate buffered saline (PBS), goat anti-rabbit secondary antibody (sp-9001, Zhongshan Golden Bridge) was added for 60 min at room temperature. Subsequently, slides were incubated with horseradish peroxidase (HRP) (sp-9001, Zhongshan Golden Bridge). The slides were lightly counterstained with hematoxylin and dehydrated. Finally, images were captured with Leica DM2000 microscope.
Sp 9001
Manufactured by Zhongshan Golden Bridge Biotechnology
Sourced in China
The SP-9001 is a high-performance laboratory centrifuge designed for a variety of applications. It features a compact, user-friendly design and can accommodate a range of sample volumes and tube sizes. The centrifuge is equipped with a digital display and intuitive control panel for easy operation.
Automatically generated - may contain errors
Lab products found in correlation
Dm2000 microscope, by Leica (1 mentions)
Ab172618, by Abcam (1 mentions)
E400 light microscope, by Nikon (1 mentions)
Ab197678, by Abcam (1 mentions)
Ab283575, by Abcam (1 mentions)
Ab92536, by Abcam (1 mentions)
Ab182981, by Abcam (1 mentions)
Ab16667, by Abcam (1 mentions)
Cy5096, by Abways (1 mentions)
Dig 3009, by Maixin Group (1 mentions)
Dab 0031, by Maixin Group (1 mentions)
Rt pcr kit, by Thermo Fisher Scientific (1 mentions)
Ab28244, by Abcam (1 mentions)
Ab16661, by Abcam (1 mentions)
Trizol rna extraction kit, by Takara Bio (1 mentions)
Hmgb1, by Thermo Fisher Scientific (1 mentions)
Sa00001 2, by Proteintech (1 mentions)
7 protocols using sp 9001
1
Immunohistochemical Analysis of CAMK1D
2
Immunohistochemical Analysis of Tumor Angiogenesis
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Briefly, paraffin-embedded tissue sections were deparaffinized, dehydrated, heated in a pressure pot for 10 min to retrieve antigens, and then treated with 0.3% H2O2. Thereafter, The sections were incubated with anti-Ki67 (1:200, ab16667, Abcam, Cambridge, England, UK), anti-CD31 (1:2000, ab182981, Abcam, Cambridge, England, UK), anti-VEGF (1:200, CY5096, Abways, Minhang District, Shanghai, China), anti-MMP9 (1:5000, ab283575, Abcam, Cambridge, England, UK), anti-MMP2 (1:200, ab92536, Abcam, Cambridge, England, UK), anti-CD66b (1:200, ab197678, Abcam, Cambridge, England, UK), and followed with secondary antibody incubation tagged with the peroxidase enzyme (1:200, SP-9001, Zhongshan Golden Bridge, Haidian, Beijing, China) for 30 min. The slides were finally visualized with 0.05% DAB and were counterstained with hematoxylin and then observed using Nikon E400 Light Microscope.
3
Histological and Immunohistochemical Analysis of Ligamentum Flavum and Synovium
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LBs and synovium specimens were fixed in 4% paraformaldehyde solution. For decalcification, LBs were immersed in a solution containing 10% ethylene diamine tetraacetic acid (EDTA) for 3 months. After a series of classic treatments for histological observations, the paraffin-embedded sections of 4 μm-thick were acquired, and disposed with haematoxylin and eosin (HE) staining.
Streptavidin-peroxidase conjugated method was applied for IHC observations, as described previously29 (link). Antigen was retrieved using pepsin (DIG-3009, Maixin, China) at 37°C for 30 min. Rabbit-originated antibodies against CD34 (1:400, ZA-0550, Zhongshan Golden Bridge Biotechnology Co., Ltd., China), human TGF-β3 (1:200, 18942-1-AP, Proteintech), and human FGF-2 (1:500, ZS-79, Zhongshan Golden Bridge Biotechnology Co., Ltd., China) were used as primary antibodies and incubated at 4°C for 18 h. The histological sections were then stained by the anti-rabbit streptavidin-peroxidase kit (SP-9001, Zhongshan Golden Bridge Biotechnology Co., Ltd., China). Finally, color development was achieved by reacting with 3, 3′-diaminobenzidine (DAB, 0031, Maixin, China). Hematoxylin was used for counterstaining.
Streptavidin-peroxidase conjugated method was applied for IHC observations, as described previously29 (link). Antigen was retrieved using pepsin (DIG-3009, Maixin, China) at 37°C for 30 min. Rabbit-originated antibodies against CD34 (1:400, ZA-0550, Zhongshan Golden Bridge Biotechnology Co., Ltd., China), human TGF-β3 (1:200, 18942-1-AP, Proteintech), and human FGF-2 (1:500, ZS-79, Zhongshan Golden Bridge Biotechnology Co., Ltd., China) were used as primary antibodies and incubated at 4°C for 18 h. The histological sections were then stained by the anti-rabbit streptavidin-peroxidase kit (SP-9001, Zhongshan Golden Bridge Biotechnology Co., Ltd., China). Finally, color development was achieved by reacting with 3, 3′-diaminobenzidine (DAB, 0031, Maixin, China). Hematoxylin was used for counterstaining.
4
ESCC Tumor Tissue Analysis Protocol
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The paraffin-embedded specimens of tumor tissues of 75 ESCC patients were collected. A total of 37 males and 38 females aged 24–78 years were included in this study, with a median age of 52 years. All the patients received operation in the Affiliated Jinling Hospital of Nanjing Medical University (Nanjing, China) from June 2009 to March 2012. The patients were pathologically diagnosed with ESCC and underwent operative treatment for the first time without receiving chemotherapy, radiotherapy and other treatments. This study was reviewed and approved by the Ethics Committee of the Affiliated Jinling Hospital of Nanjing Medical University, and all patients or their families signed the informed consent.
Tools included in this study: DNA extraction kits and reverse transcription-polymerase chain reaction (RT-PCR) kits (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA); restriction endonuclease Aci I (New England Biolabs, Wilbury WayHitchin, Herts, UK); rabbit monoclonal Bax antibody (1:100; cat. no. ab32503, Abcam; Cambridge, MA, USA); immunohistochemical staining kits SP-9001 (Zhongshan Goldenbridge Biotechnology, Beijing, China); primer synthesis (Takara, Dalian, China).
Tools included in this study: DNA extraction kits and reverse transcription-polymerase chain reaction (RT-PCR) kits (Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA); restriction endonuclease Aci I (New England Biolabs, Wilbury WayHitchin, Herts, UK); rabbit monoclonal Bax antibody (1:100; cat. no. ab32503, Abcam; Cambridge, MA, USA); immunohistochemical staining kits SP-9001 (Zhongshan Goldenbridge Biotechnology, Beijing, China); primer synthesis (Takara, Dalian, China).
5
Uterine Tissue Histological Analysis
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For hematoxylin and eosin staining, the uterus was fixed overnight in 4% paraformaldehyde. Then, it was embedded in paraffin, sectioned, and stained.
For immunohistochemistry analysis, the paraffin blocks were sectioned and blocked with a blocking buffer. Immunostaining was performed using the primary antibodies estrogen receptor (ER; anti-rabbit, ab3206, Abcam), progesterone receptor (PR; anti-rabbit, ab16661, Abcam), and fibroblast activation protein (FAP; anti-rabbit, ab28244, Abcam) and visualized using goat anti-rabbit antibody (SP-9001, Beijing Zhong Shan Golden Bridge Biotechnology, China) and 3,3′-diaminobenzidine stain.
For immunohistochemistry analysis, the paraffin blocks were sectioned and blocked with a blocking buffer. Immunostaining was performed using the primary antibodies estrogen receptor (ER; anti-rabbit, ab3206, Abcam), progesterone receptor (PR; anti-rabbit, ab16661, Abcam), and fibroblast activation protein (FAP; anti-rabbit, ab28244, Abcam) and visualized using goat anti-rabbit antibody (SP-9001, Beijing Zhong Shan Golden Bridge Biotechnology, China) and 3,3′-diaminobenzidine stain.
6
Tumor and Normal Tissue Samples for STS
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Frozen and paraffin specimens of tumor tissues and corresponding para-carcinoma normal tissues were collected from 64 STS patients treated in the General Hospital of Chinese People's Liberation Army (Beijing, China), from January 2007 to January 2012. Patients were aged 11–72 years, including 35 males and 29 females, all of them were pathologically diagnosed with STS, and they had complete clinical data and received surgical treatment in the hospital.
This study was reviewed and approved by the Clinical Ethics Committee of General Hospital of Chinese People's Liberation Army, and all patients or their families signed an informed consent.
TRIzol RNA extraction kit (Takara, Dalian, China), primer synthesis, RT kits and quantitative SYBR-Green PCR kits (Ambion; Thermo Fisher Scientific, Inc., Waltham, MA, USA), primary rabbit anti-human p16 and nm23-H1 antibodies (1:100; cat. nos. 10883-1-AP and 11086-2-AP, respectively; Proteintech Group, Inc., Wuhan, China), immunohistochemical staining kit SP-9001 (Beijing Zhongshan Goldenbridge Biotechnology Co., Ltd., Beijing, China).
This study was reviewed and approved by the Clinical Ethics Committee of General Hospital of Chinese People's Liberation Army, and all patients or their families signed an informed consent.
TRIzol RNA extraction kit (Takara, Dalian, China), primer synthesis, RT kits and quantitative SYBR-Green PCR kits (Ambion; Thermo Fisher Scientific, Inc., Waltham, MA, USA), primary rabbit anti-human p16 and nm23-H1 antibodies (1:100; cat. nos. 10883-1-AP and 11086-2-AP, respectively; Proteintech Group, Inc., Wuhan, China), immunohistochemical staining kit SP-9001 (Beijing Zhongshan Goldenbridge Biotechnology Co., Ltd., Beijing, China).
7
Ovarian Cancer Tissue Analysis
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In the present study, tumor tissues and the corresponding para-carcinoma normal tissues of 60 ovarian cancer patients admitted into the Department of Surgery in Dongying Hospital (Dongying, China) from June, 2012 to June, 2015, were selected. The patients were definitively diagnosed with ovarian cancer by clinical pathology and did not receive chemotherapy and surgical therapy. Patients were aged 24–78 years, and the median age was 52 years. This study was approved by the Clinical Ethics Committee of Dongying People's Hospital. At the same time, all the patients enrolled signed the informed consent form.
RNA extraction, reverse transcription, and RT-qPCR kit (all from Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA), HMGB1, BRCA1, P62, GAPDH antibodies (1:800; cat. nos. 10829-1-AP, 22362-1-AP, 18420-1-AP, 10494-1-AP) and HRP-labeled secondary antibodies (1:1000; cat. no. SA00001-2) (all from Proteintech Biotechnology Co., Ltd.; Wuhan Sanying Biotechnology, Wuhan, China); immunohistochemical staining kit SP-9001 (Beijing Zhongshan Goldenbridge Biotechnology Co., Ltd.; OriGene Technologies, Inc., Beijing, China), primer synthesis (Takara Biotechnology Co., Ltd., Dalian, China) were used in the present study.
RNA extraction, reverse transcription, and RT-qPCR kit (all from Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA), HMGB1, BRCA1, P62, GAPDH antibodies (1:800; cat. nos. 10829-1-AP, 22362-1-AP, 18420-1-AP, 10494-1-AP) and HRP-labeled secondary antibodies (1:1000; cat. no. SA00001-2) (all from Proteintech Biotechnology Co., Ltd.; Wuhan Sanying Biotechnology, Wuhan, China); immunohistochemical staining kit SP-9001 (Beijing Zhongshan Goldenbridge Biotechnology Co., Ltd.; OriGene Technologies, Inc., Beijing, China), primer synthesis (Takara Biotechnology Co., Ltd., Dalian, China) were used in the present study.
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