Xenograft tumors were fixed with 10% formaldehyde and embedded in paraffin, followed by being sliced, dewaxed, and hydrated. Antigen retrieval was performed using 0.01 M citrate buffer. H2O2 solution (3%) was applied to incubate sections for 15 minutes at room temperature. Goat serum (5%) was added onto sections for 30 minutes incubation at room temperature. Thereafter, sections were sequentially subjected to incubation with primary antibody (1:100, mouse anti-Ki67 and anti-PTEN, Geneseed Biotech, Guangzhou, China) overnight at 4°C and with HRP labeled secondary antibody (1:300, Boster, Wuhan, China) for 30 minutes at room temperature. After being stained with diaminobenzidine (DAB) and hematoxylin, sections were sealed with neutral resin. The positive signals of Ki67 and PTEN were observed under a microscope (Olympus, Tokyo, Japan).
Hrp labeled secondary antibody
Manufactured by Boster Bio
Sourced in China
The HRP labeled secondary antibody is a laboratory reagent used in various immunoassay techniques. It consists of a secondary antibody conjugated with the enzyme horseradish peroxidase (HRP). The HRP label allows for the detection and amplification of the target antigen-antibody interaction through a colorimetric or chemiluminescent reaction.
Automatically generated - may contain errors
Lab products found in correlation
Thalidomide, by Merck Group (1 mentions)
Anti β actin, by Boster Bio (1 mentions)
P gsk3β, by Cell Signaling Technology (1 mentions)
Mmp 2, by R&D Systems (1 mentions)
Tgf β1 receptor kinase inhibitor sb431542, by Selleck Chemicals (1 mentions)
Jnk inhibitor sp600125, by Merck Group (1 mentions)
Tgf β1, by R&D Systems (1 mentions)
Mmp 9, by R&D Systems (1 mentions)
Immunohistochemistry kit, by Boster Bio (1 mentions)
Real time pcr kit, by Takara Bio (1 mentions)
Anti tlr4, by Abcam (1 mentions)
Anti myd88, by Abcam (1 mentions)
Anti trif, by Abcam (1 mentions)
Anti traf6, by Abcam (1 mentions)
Rnaios, by Takara Bio (1 mentions)
Trizol reagent, by Takara Bio (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
4 protocols using hrp labeled secondary antibody
1
Immunohistochemical Analysis of Xenograft Tumors
2
Evaluating Cell Proliferation via Ki-67 Immunostaining
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The tumor tissues were fixed with 4% paraformaldehyde, embedded in paraffin, and cut into 5-μm-thick sections. Then the paraffin sections were incubated with primary antibodies against Ki-67(1:100, Geneseed Biotech, Guangzhou, China) overnight at 4°C, followed with HRP labeled secondary antibody (1:300, Boster, Wuhan, China) for 30 minutes at room temperature. After being stained with diaminobenzidine (DAB) and hematoxylin, sections were sealed with neutral resin. Finally, the Immunostaining images were observed with a microscope (Olympus, Tokyo, Japan).
3
Signaling Pathways in TGF-β1 Regulation
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TGF-β1 was purchased from R&D Systems (Minneapolis, MN, USA). TGF-β1 receptor kinase inhibitor (SB431542) was purchased from Selleck (Houston, TX, USA). PI3K/Akt inhibitor (LY294002) was purchased from Cell Signaling Technology (Beverly, MA, USA). JNK inhibitor (SP600125) and ERK inhibitor (PD098059) were purchased from Millipore Co. (Billerica, MA, USA). Thalidomide was purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-rat p38 (ab7952), p-p38 (ab4822), Smad2/3 (ab63672) and p-Smad2/3 (ab63399) were purchased from Abcam (Shanghai, China). Anti-rat ERK (BS1121) and E-Cadherin (BS1097) were purchased from Bioworld (Minneapolis, USA). Anti-rat p-ERK (#4370), AKT (#9272), p-AKT (#2965s), GSK3β (#9315) and p-GSK3β (#9336) were purchased from Cell Signaling (Beverly, MA, USA). Anti-rat JNK (sc-571), p-JNK (sc-81502) and α-SMA (sc-71626) were purchased from Santa Cruz (CA, USA). Anti-rat vimentin (3634–100) was purchased from Biovision (Mountain View, CA, USA). Anti-rat ZO-1 was purchased from Invitrogen (Carlsbad, CA, USA). Anti-β-actin and HRP-labeled secondary antibodies were purchased from Boster (Wuhan, Hubei, China). Commercial ELISA kits for matrix metalloproteinase (MMP)-2 and MMP-9 were purchased from R&D Systems (Shanghai, China). ELISA kit for collagen type IV was purchased from BlueGene Biotech (Shanghai, China).
4
Molecular Pathway Analysis of TLR4-Mediated Inflammation
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Trizol reagent, RNAios and Real-time PCR kit were obtained from TaKaRa Company (Dalian, China). Anti- MyD88, anti-TRAF6, anti-TRIF and anti-TLR4 antibodies were offered from Abcam Biotechnology (MA, USA). LPS was taken from Sigma-Aldrich (MO, USA). The immunohistochemistry kits and HRP-labeled secondary antibodies were bought from Boster Biotechnology (Wuhan, China). The IL-1β and IL-6 ELISA kit were purchased from Dkewei Biotechnology (Beijing, China). Corilagin was received from China National institutes for food and drug control.
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