The protein level of KDM5A, p16, and Fbxo22 was detected using streptavidin peroxidase labeled by immunohistochemical peroxidase. Paraffin-embedded specimens of breast cancer tissues were serially sectioned (5 μm thickness) and immune-stained with primary antibodies of rabbit anti-KDM5A (1:1000, ab78322, Abcam, UK), rabbit anti-p16 (1:100, A0262, ABclonal), and rabbit anti-Fbxo22 (13,606–1-AP, Protein-tech, Wuhan, China) overnight at 4 °C. The samples were incubated at 37 °C for 20 min with biotin-labeled goat anti-rabbit secondary antibody (BA1003, Boster), followed by incubation of 50 μL of streptomyces anti-biotin–peroxidase solution for 10 min at ambient temperature. Color development was completed with 3,3′-diaminobenzidine and microscopic observation for the specimens was performed with IgG used as a NC. Protein-positive cells were identified by the brownish-yellow color of normal positive cells, and positive staining was statistically analyzed using ImageJ.
Ba1003
Manufactured by Boster Bio
Sourced in China, United States
BA1003 is a laboratory equipment item designed for use in scientific and research settings. It is a versatile piece of equipment that serves a core function, but a detailed description cannot be provided while maintaining an unbiased and purely factual approach without extrapolation or interpretation.
Automatically generated - may contain errors
Lab products found in correlation
Ab78322, by Abcam (1 mentions)
A0262, by ABclonal (1 mentions)
Rabbit antibodies, by Abcam (1 mentions)
Dm 4000b photomicroscope, by Leica (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Vectastain elite abc reagent, by Vector Laboratories (1 mentions)
3 3 diaminobenzidine (dab), by Merck Group (1 mentions)
Gelatin coated slides, by Merck Group (1 mentions)
Ab60134, by Abcam (1 mentions)
Ab78007, by Abcam (1 mentions)
Ab46154, by Abcam (1 mentions)
Ab52915, by Abcam (1 mentions)
Df6701, by Affinity Biosciences (1 mentions)
G1006, by Wuhan Servicebio Technology (1 mentions)
Bm5318, by Boster Bio (1 mentions)
Ab182795, by Abcam (1 mentions)
Ab39012, by Abcam (1 mentions)
Ab78066, by Abcam (1 mentions)
Ab84036, by Abcam (1 mentions)
Ab55735, by Abcam (1 mentions)
11 protocols using ba1003
1
Immunohistochemical Analysis of KDM5A, p16, and Fbxo22 in Breast Cancer
2
Immunohistochemical Analysis of Stem Cell Markers
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Paraffin tissue samples were sectioned at the thickness of 5 μm, dehydrated, treated with 3% hydrogen peroxide at room temperature for 10 min and blocked with normal goat serum for 10 min. Next, the sections were immunostained with rabbit antibodies (Abcam) against ALKBH5 (1:2000, ab195377), UBE2C (1:500, ab252940), p53 (1:100, ab32389), Ki67 (1:200, ab15580), NANOG (1:200, ab109250), OCT4 (1:250, ab200834), and SOX2 (1:100, ab92494) at 4 °C overnight. The next day, the sections were incubated with biotin-labeled goat anti-rabbit secondary antibody (1:200, BA1003, Boster Biological Technology Co. Ltd., Wuhan, Hubei, China) at 37 °C for 20 min and then with 50 μL streptavidin biotin peroxidase complex at room temperature for 10 min. The sections were exposed to DAB substrate, which was followed by hematoxylin counterstaining and dehydration treatment. The staining images were obtained using a microscope. The normal positive cells were brownish yellow, and the positive staining was statistically analyzed by ImageJ software.
3
IHC Analysis of RZR/ROR and VEGF
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Mice were sacrificed using the cervical dislocation method, and tumors were excised 7 days following MLT treatment. The excised tumors were cut into 5-µm sections, and transferred to gelatin-coated slides (Sigma-Aldrich). Following a 30-min incubation in phosphate-buffered saline (PBS) containing 0.3% Triton X-100 (Sigma-Aldrich), the sections were incubated with a primary antibody against RZR/ROR receptor [rabbit anti-RZR/RORα polyclonal antibody (1:500; catalog number ab60134; Abcam, Cambridge, MA, USA), rabbit anti-RZR/RORβ polyclonal antibody (1:500; catalog number ab78007; Abcam) or rabbit anti-RZR/RORγ polyclonal antibody (1:500; catalog number ab188756; Abcam) or vascular endothelial growth factor (VEGF) (rabbit anti-VEGF polyclonal antibody (1:200; catalog number ab46154; Abcam) at 37°C overnight. After washing with 1X-PBS three times, the sections were incubated with a polyclonal goat anti-rabbit biotinylated antibody (1:100; catalog number BA1003; Boster Biological Technology Co., Pleasanton, CA, USA) for 1 h, and subsequently incubated in VECTASTAIN® Elite ABC Reagent (Vector Laboratories, Inc., Burlingame, CA, USA). Subsequently, the sections were stained with 3,3′-diaminobenzidine (DAB; Sigma-Aldrich). Images were taken with a Leica DM4000B photo microscope (Leica Microsystems GmbH, Wetzlar, Germany; magnification, ×400).
4
Penile Tissue Immunohistochemistry for Oxidative Stress
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IHC staining of penile tissue sections was performed using antibodies against MMP3 (ab52915, Abcam, London, UK), CDH1 (20874‐1‐AP, Proteintech, Wuhan, China), CD71 (10084‐2‐AP, Proteintech, Wuhan, China), ZIP8 (20459‐1‐AP, Proteintech, Wuhan, China), GPX4 (DF6701, Affinity Biosciences, Changzhou, China), SLC7A11 (BM5318, Boster, Wuhan, China), and ACSL4 (ab240135, Abcam, London, UK). A biotin‐conjugated secondary antibody (BA1003, Boster, Wuhan, China) was then used to incubate the sections. Based on the manufacturer's recommendations, Masson's trichrome staining (G1006, Servicebio, Wuhan, China) was used to evaluate changes in tissue structure. It was also possible to quantify the ratio of smooth muscle to collagen in the corpus cavernosum.
5
Immunohistochemical Analysis of Iron Metabolism and Cartilage Degradation
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From these decalcified samples, 5 μm sagittal sections were cut from the medial knee compartment and were deparaffinized, antigen retrieved, incubated with corresponding primary antibodies TFR1 (ab84036, Abcam, dilution 1:3000), DMT1 (ab55735, Abcam, dilution 1:1000), FPN(ab78066, Abcam, dilution 1:100), MMP13 (ab39012, Abcam, dilution 1:1000), ADAMTS5 (ab182795, Abcam, dilution 1:1000), and then incubated with biotinylated goat anti-rabbit (BA1003, Boster, China, dilution 1:100) secondary antibodies. Sections were colored with DAB and counterstained with hematoxylin. The images of immunohistochemical staining were analyzed using the software Image-Pro Plus. To quantify immuno-positive cells, at least five random fields in the cartilage were selected and the ratio of immune positive cells to total cells were calculated.
6
Immunohistochemical Analysis of Femoral Angiogenesis
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After CT scanning, femurs were decalcified with 10% EDTA solution for 3 weeks and embedded in paraffin wax. The femoral heads were sectioned at 5-μm thickness in the coronal plane. Some of these sections were stained with hematoxylin and eosin (H&E) to evaluate the trabecular structure, while the others were deparaffinized; antigen retrieved; incubated with anti-CD31, anti-VEGF, and anti-VEGFR2 primary antibodies; and then incubated with the corresponding biotinylated goat anti-rabbit (Boster, China, BA1003) and goat anti-mouse (Boster, China, BA1001) secondary antibodies. Sections were colored with DAB and counterstained with hematoxylin. The images of immunohistochemical staining were analyzed using the software Image-Pro Plus. At least five random fields in the femoral head were selected, and the positive staining was quantified based on the integrated option density (IOD) of target proteins. The corresponding area was also measured, and the mean density was defined as the ratio of integrated optical density to the corresponding area.
7
Immunohistochemical Analysis of NGAL Expression
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Following deparaffinization, antigen retrieval was performed in citrate buffer. Non-specific binding was blocked with 0.3% H2O2 for 10 min and, after blocking for 30 min with 10% goat serum at room temperature, anti-NGAL antibody (1:150 in PBS; PB0641; Boster Biological Technology, Ltd., Wuhan, China) was applied overnight at 4°C. Samples were washed in buffer and incubated with the goat anti-rabbit IgG-Biotin secondary antibody (1:200 in PBS; BA1003; Boster Biological Technology, Ltd.) for 30 min, followed by the diaminobenzidine-peroxidase reaction and counterstaining with hematoxylin. Negative control sections were stained under identical conditions by omitting the primary antibody instead of PBS. Stained sections were subsequently visualized using light microscopy at ×200 magnification and the integrated optical density (IOD) was measured using Image-Pro Plus 6.0 (Media Cybernetics, Inc., Rockville, MD, USA).
8
Immunohistochemical Analysis of NLRP3, Caspase-3, and LC3
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Immunohistochemical staining was performed as follows: sections were incubated with NLRP3 (bs-10021R; Bioss Antibodies, Woburn, MA, USA), caspase-3 (bs-20364R; Bioss Antibodies), and LC3 (14600-1-AP, Proteintech, Inc., Rosemont, IL, USA) antibodies for 12 hours at 4 °C. Following a rinse, sections were exposed to biotin-conjugated goat anti-rabbit immunoglobin G (IgG) secondary antibody (BA1003, Boster Bio, Pleasanton, CA, USA), and subsequently, streptavidin-horseradish peroxidase (HRP) (bs-0437P, Bioss Antibodies) conjugation for 2 hours at room temperature (RT). Finally, the sections were stained with DAB (ZLI-9019; ZSGB-BIO, Beijing, China) and hematoxylin before observation under a light microscope.
9
Immunohistochemical Analysis of P2X3 Receptor
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The DRG tissues were fixed with 4% paraformaldehyde for 12 h before conventional dehydration and then cut into 4 um sections after paraffin embedding. The sections were routinely deparaffinized, rehydrated, and incubated with 0.3% H2O2 for 20 min, followed by washing (5 min ×2) with distilled water. The slides were then blocked in normal goat serum at room temperature for 20 min. Next, drops of primary antibody were added, and the slides were incubated at 37°C for 2 h (anti-P2X3 primary antibody, Abcam, ab10269, 1 : 1000 dilution), followed by washing (5 min ×3) with 0.01 M PBS. The secondary antibody (goat anti-rabbit IgG, BOSTER Biological Technology, BA1003, 1 : 100 dilution) was then added and incubated at room temperature for 30 min, followed by washing (5 min ×3) with 0.01 M PBS and staining with diamino-benzidine (DAB) and hematoxylin. Image acquisition was conducted using an Olympus-BX53 microscope, and Image-Pro PLUS software was used for the data acquisition.
10
Rbpjl Immunohistochemical Staining in Pancreatic Tissue
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Pancreatic tissues were sectioned at the thickness of 5 μm, dehydrated, treated with 3% hydrogen peroxide, and then subjected to peroxidase blocking with normal goat serum. Immunohistochemical staining was subsequently performed by incubating the tissue sections with primary antibody to Rbpjl (dilution ratio of 1:100, 14,613–1-AP, Proteintech) overnight at 4 °C. The following day, the tissues sections were incubated with biotin-labeled goat anti-rabbit secondary antibody (dilution ratio of 1:200, BA1003, Boster Biological Technology Co. Ltd., Wuhan, Hubei, China) for 20 min at 37 °C. Afterwards, the sections were exposed to DAB substrate, followed by hematoxylin counterstaining and standard dehydration treatment. Staining images were obtained using a microscope. Finally, the ImageJ software was adopted for statistical analysis of positive staining. Five different fields of view were randomly selected to count the number of positive cells, and the total number of cells counted was more than 100. The proportion of positive cells = (number of positive cells/total cells) × 100% [40 (link)].
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