After deparaffinization, rehydration and antigen retrieval, the sections were incubated with anti-MPO antibody (1:100; ab9535; Abcam, Cambridge, UK), anti-IL-1β antibody (1:100; SC-7884; Santa Cruz Biotechnology, CA, USA), and anti-IL-6 antibody (1:100; SC-1265-R; Santa Cruz, CA, USA) overnight at 4 °C7 (link),8 (link). After incubation with goat anti-rabbit secondary antibody (Boster, Wuhan, China), the samples were visualized with a 3,3-diaminobenzidine (DAB) kit (Boster, Wuhan, China). The mounted sections were observed and photographed under a microscope at 200 × magnification (DM2500; Leica, Solms, Germany).
3 3 diaminobenzidine dab kit
Manufactured by Boster Bio
Sourced in China
The 3,3-diaminobenzidine (DAB) kit is a laboratory reagent used for the detection and visualization of specific protein targets in immunohistochemistry (IHC) and other applications. The kit contains the necessary components for the chromogenic detection of labeled antibodies or other detection probes.
Automatically generated - may contain errors
Lab products found in correlation
Dm2500, by Leica (5 mentions)
Goat anti rabbit secondary antibody, by Boster Bio (4 mentions)
Ab9535, by Abcam (2 mentions)
Sc 7884, by Santa Cruz Biotechnology (2 mentions)
Sc 1265 r, by Santa Cruz Biotechnology (2 mentions)
Sabc pod rabbit igg kit, by Boster Bio (1 mentions)
Mpo antibody, by Abcam (1 mentions)
Anti p bad, by Cell Signaling Technology (1 mentions)
Anti p akt, by Cell Signaling Technology (1 mentions)
Sabc pod kit, by Boster Bio (1 mentions)
Nb500 170, by Santa Cruz Biotechnology (1 mentions)
Nbp2 13740, by Proteintech (1 mentions)
He staining kit, by Solarbio (1 mentions)
Biotinylated anti rabbit secondary antibodies, by Boster Bio (1 mentions)
7 protocols using 3 3 diaminobenzidine dab kit
1
Immunohistochemical Analysis of MPO, IL-1β, and IL-6
2
Establishing OSCC PDX Models and Evaluating Anlotinib
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The OSCC PDX models were established as previously described [16 (link)]. In brief, eight patients’ samples were used to establish the PDX models and the PDX mice were treated with 3 mg/kg anlotinib or normal saline (oral administration, once a day). The tumor growth inhibition (TGI) rate was calculated as (1-(Tn-T0)/(Cn-C0)) × 100% and presented in our previous study [16 (link)].
Tissue slides from PDX models were routinely deparaffinized and rehydrated. Following endogenous peroxidase quenching and antigen retrieval, the slides were blocked with 5% BSA for 30 min, followed by incubation with primary antibodies against METTL3 (Proteintech, 15,073–1-AP), FGFR3 (Signalway Antibody, 33,373) and p-FGFR3 (Affinity Biosciences, AF8439) overnight at 4℃. Then secondary antibodies and SABC were applied with SABC-POD (rabbit IgG) kit (BOSTER, Wuhan, China). The slides were stained using 3, 3’-diaminobenzidine (DAB) kit (BOSTER) and then counterstained with hematoxylin.
Tissue slides from PDX models were routinely deparaffinized and rehydrated. Following endogenous peroxidase quenching and antigen retrieval, the slides were blocked with 5% BSA for 30 min, followed by incubation with primary antibodies against METTL3 (Proteintech, 15,073–1-AP), FGFR3 (Signalway Antibody, 33,373) and p-FGFR3 (Affinity Biosciences, AF8439) overnight at 4℃. Then secondary antibodies and SABC were applied with SABC-POD (rabbit IgG) kit (BOSTER, Wuhan, China). The slides were stained using 3, 3’-diaminobenzidine (DAB) kit (BOSTER) and then counterstained with hematoxylin.
3
Immunohistochemical Analysis of Myeloperoxidase
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Paraffin-embedded tissues (5-μm-thick slices) were examined by IHC and IF staining. Some sections were incubated with anti-myeloperoxidase (MPO) antibodies (Abcam, Cambridge, UK) overnight at 4°C. Then, they were incubated with goat anti-rabbit secondary antibody (Boster, Wuhan, China), and visualised with a 3,3-diaminobenzidine (DAB) kit (Boster, Wuhan, China). Finally, the mounted sections were observed and photographed under a microscope at 200× magnification (DM2500, Leica, Solms, Germany).
4
Immunohistochemical Analysis of p-Akt and p-Bad
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Paraffin-embedded tissues were cut into 5-μm-thick slices for IHC examination. The sections were incubated with anti-p-Akt (1:200) and anti-p-Bad (1:200) antibodies (both from Cell Signaling Technology, Boston, MA, USA) overnight at 4 °C. Then, they were incubated with goat anti-rabbit secondary antibody (Boster, Wuhan, China), and visualized with a 3,3-diaminobenzidine (DAB) kit (Boster, Wuhan, China). Finally, the mounted sections were observed and photographed under a microscope at 200× magnification (DM2500, Leica, Solms, Germany).
5
Immunohistochemical Profiling of Tumor Markers
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All specimens collected were routinely embedded in paraffin, and the tissue slides were deparaffinized and rehydrated. H&E staining was performed with an H&E staining kit (Solarbio, G1120-100). For IHC staining, after incubation with 3% H2O2, antigen retrieval, and blocking, sections were incubated with primary anti-PCIF1 (Novus, NBP2-13740), anti-CTBP2 (Proteintech, 10346), anti-TET2 (Abcam, ab94580), anti-Ki67 (Novus, NB500-170), and anti-PCK (pan-cytokeratin; Santa Cruz, sc-8018) overnight at 4°C. After washing, secondary antibodies and streptavidin-biotin complex (SABC) were applied with an SABC-POD kit (BOSTER, SA1022). Then the slides were stained with a 3,3′-diaminobenzidine (DAB) kit (BOSTER) and counterstained with hematoxylin. For the IHC analysis, the IHC score was calculated by multiplying the proportion of positively stained cells at each intensity level by their respective intensity score (intensity scores ranging from 0 to 3). The final score ranges from 0 to 300. For Ki67, the percentage of positively stained cells was calculated. The anti-PCK antibody (Santa Cruz Biotechnology, sc-8018) was used for IHC staining of metastatic cells in cervical lymph nodes.
6
Amyloid-β Immunohistochemistry Protocol
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The sections were incubated with 0.3% H2O2 in methanol for 10 min to inhibit endogenous peroxidases. After three washed with PBS, the sections were blocked with 10% normal goat serum for 20 min at room temperature, and then incubated with primary antibody anti-Aβ1-42 (1 : 200, 14974, CST, USA) overnight at 4 °C. After washing, the sections were incubated with biotinylated anti-rabbit secondary antibodies (Boster, China) in PBS for 30 min at 37 °C. Then, the sections were incubated with avidinbiotin peroxidase solution (SABC kit, Boster, China) and colorized with a 3,3-diaminobenzidine (DAB) kit (Boster, China). The sections were observed under a light microscope (Leica DM2500, Germany).
7
Renal Inflammation Mediator IHC Analysis
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IHC staining was performed for evaluation on distribution of in ammation mediators in the renal tissues.
After depara nization, rehydration and antigen retrieval, the sections were incubated with anti-MPO antibody (1:100, ab9535, Abcam, Cambridge, UK), anti-IL-1β antibody (1:100, SC-7884, Santa Cruz Biotechnology, CA, USA), and anti-IL-6 antibody (1:100, SC-1265-R, Santa Cruz, CA, USA) overnight at 4 ºC.
After incubating goat anti-rabbit secondary antibody (Boster, Wuhan, China), the samples were visualized with a 3,3-diaminobenzidine (DAB) kit (Boster, Wuhan, China). The mounted sections were observed and photographed under a microscope at 200× magni cation (DM2500, Leica, Solms, Germany).
After depara nization, rehydration and antigen retrieval, the sections were incubated with anti-MPO antibody (1:100, ab9535, Abcam, Cambridge, UK), anti-IL-1β antibody (1:100, SC-7884, Santa Cruz Biotechnology, CA, USA), and anti-IL-6 antibody (1:100, SC-1265-R, Santa Cruz, CA, USA) overnight at 4 ºC.
After incubating goat anti-rabbit secondary antibody (Boster, Wuhan, China), the samples were visualized with a 3,3-diaminobenzidine (DAB) kit (Boster, Wuhan, China). The mounted sections were observed and photographed under a microscope at 200× magni cation (DM2500, Leica, Solms, Germany).
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