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Ki 67 h 300

Manufactured by Santa Cruz Biotechnology
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Ki-67 (H-300) is a lab equipment product that serves as an antibody. It is commonly used in research applications to detect the presence and distribution of the Ki-67 protein, which is a marker of cellular proliferation.

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Lab products found in correlation

F4 80, by Bio-Rad (2 mentions) Biotinylated horse anti mouse igg, by Vector Laboratories (2 mentions) Biotinylated horse anti rabbit igg, by Vector Laboratories (2 mentions) 3 3 diaminobenzidine (dab), by Vector Laboratories (2 mentions) Novocastra streptavidin hrp, by Leica (2 mentions) Aec high sensitivity substrate chromogen, by Agilent Technologies (2 mentions) Cdk4 c 22, by Santa Cruz Biotechnology (2 mentions) Vectastain elite abc kit, by Vector Laboratories (2 mentions) Nitrocellulose membrane, by Cytiva (1 mentions) Quantity one analysis system, by Bio-Rad (1 mentions) Pcna fl 261, by Santa Cruz Biotechnology (1 mentions) β actin c4, by Santa Cruz Biotechnology (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Alexa fluor 488 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) E cadherin, by BD (1 mentions)

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H-300 (126 citations)

5 protocols using ki 67 h 300

1

Western Blot Analysis of Cell Signaling Proteins

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The cells were lysed in radioimmunoprecipitation assay (RIPA) lysis buffer (Beyotime Biotech, Jiangsu, China), and centrifuged at 12000× g for 45 min at 4°C. After harvesting the supernatant, protein concentration was determined using the BCA assay kit KeyGEN (Beyotime Biotech). Samples containing 60 μg of protein were resolved on 12% SDS-PAGE electrophoresis and transferred to nitrocellulose membranes (Whatman International Ltd., Germany). After blocking for 1 h, the membranes were incubated with the following primary antibodies overnight at 4°C: Ki-67 (H-300), PCNA (FL-261), cyclin E (M20), cyclin D1 (H-295), p27 (C-19), p21 (C-19), CDK2 (D-12), CDK4 (C-22), CDK6 (C-21), p-ERK 1/2 (E-4), ERK 1/2 (H-72), p-JNK (G-7), JNK (D-2), p-AKT/Thr308 (p-AKT), AKT, and β-actin (C4): dilution 1: 200 (Santa Cruz Biotechnology) antibody. After being washed 3 times with TBST buffer, the membranes were incubated with corresponding horseradish peroxidase-conjugated secondary antibodies for 1.5 h at room temperature and analyzed using the Quantity One analysis system (Bio-Rad, Hercules, CA, USA).
Wang B., Zhou X., Wang Y, & Li R. (2017). Trifluoperazine Inhibits Mesangial Cell Proliferation by Arresting Cell Cycle-Dependent Mechanisms. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 23, 3461-3469.
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2

E-cadherin and Ki-67 Immunofluorescence

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Co-culture plates were fixed on day 6 with 4% formaldehyde for 10 minutes followed by permeabilization with 0.1% Triton X-100 for 20 minutes. After blocking with 2% BSA for 1hour, the cells were incubated with primary antibody against E-cadherin (Cat# 610182, BD Biosciences) at 1:100 dilution and Ki-67 (H300) (Cat# sc-15402 Santa Cruz, Dallas, Texas) at 1:50 dilution at 4°C overnight and then with Alexa Fluor 488-conjugated secondary antibodies (Life Technologies) at room temperature for 1 hour. DAPI was applied to stain the nucleus for 5 minutes.
Furukawa M., Wheeler S., Clark A.M, & Wells A. (2015). Lung Epithelial Cells Induce Both Phenotype Alteration and Senescence in Breast Cancer Cells. PLoS ONE, 10(1), e0118060.
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3

Measuring Tumor Cell Proliferation and Apoptosis

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The tumor tissues derived from subcutaneously injected KHOS/NP were harvested and subjected to IHC for Ki-67: H-300 (Santa Cruz Biotechnology) and cleaved caspase-3: D3E9 (Cell Signaling).
Iyer S.V., Parrales A., Begani P., Narkar A., Adhikari A.S., Martinez L.A, & Iwakuma T. (2015). Allele-specific silencing of mutant p53 attenuates dominant-negative and gain-of-function activities. Oncotarget, 7(5), 5401-5415.
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4

Immunohistochemical Analysis of Mouse Tissues

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Mouse tissues were washed, fixed in 4% paraformaldehyde, dehydrated, embedded in paraffin and sectioned. Feet tissue was additionally decalcified with 15% EDTA pH 7.2. Primary antibodies used for immunohistochemical stainings were CDK4 (C-22) from Santa Cruz, p-S6 S240/244 (rabbit polyclonal) and cleaved caspase 3 (5A1E) were purchased from CST. Ki-67 (H-300) was from Santa Cruz or from Abcam (ab66155), Mac-2 (M3/38) was obtained from Cederlane and MPO was from Dako. Biotinylated horse anti-mouse IgG or biotinylated horse anti-rabbit IgG were from Vector Laboratories. Novocastra streptavidin-HRP (Leica) and AEC-high sensitivity substrate chromogen (Dako) were used for detection of primary antibodies. Alternatively VECTASTAIN Elite ABC Kit and DAB (Vector Laboratories) were utilized. For immunofluorescent staining of paraffin slides, primary antibodies used were p-S6 S240/244 from CST, Mac-2 (M3/38) either from Cederlane or directly labelled from Biolegend and F4/80 (CI:A3-1) from Serotec. Species-matched secondary antibodies (A488 and A546) were purchased from Invitrogen. Some images were quantitatively analyzed with ImageJ. Percentage of positive staining was determined by calculating the ratio between stained area and total tissue area within a predefined threshold. All pixels in the image with values below the threshold were counted.
Linke M., Pham H.T., Katholnig K., Schnöller T., Miller A., Demel F., Schütz B., Rosner M., Kovacic B., Sukhbaatar N., Niederreiter B., Blüml S., Kuess P., Sexl V., Müller M., Mikula M., Weckwerth W., Haschemi A., Susani M., Hengstschläger M., Gambello M.J, & Weichhart T. (2017). Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression. Nature immunology, 18(3), 293-302.
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5

Immunohistochemical Analysis of Mouse Tissues

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Mouse tissues were washed, fixed in 4% paraformaldehyde, dehydrated, embedded in paraffin and sectioned. Feet tissue was additionally decalcified with 15% EDTA pH 7.2. Primary antibodies used for immunohistochemical stainings were CDK4 (C-22) from Santa Cruz, p-S6 S240/244 (rabbit polyclonal) and cleaved caspase 3 (5A1E) were purchased from CST. Ki-67 (H-300) was from Santa Cruz or from Abcam (ab66155), Mac-2 (M3/38) was obtained from Cederlane and MPO was from Dako. Biotinylated horse anti-mouse IgG or biotinylated horse anti-rabbit IgG were from Vector Laboratories. Novocastra streptavidin-HRP (Leica) and AEC-high sensitivity substrate chromogen (Dako) were used for detection of primary antibodies. Alternatively VECTASTAIN Elite ABC Kit and DAB (Vector Laboratories) were utilized. For immunofluorescent staining of paraffin slides, primary antibodies used were p-S6 S240/244 from CST, Mac-2 (M3/38) either from Cederlane or directly labelled from Biolegend and F4/80 (CI:A3-1) from Serotec. Species-matched secondary antibodies (A488 and A546) were purchased from Invitrogen. Some images were quantitatively analyzed with ImageJ. Percentage of positive staining was determined by calculating the ratio between stained area and total tissue area within a predefined threshold. All pixels in the image with values below the threshold were counted.
Linke M., Pham H.T., Katholnig K., Schnöller T., Miller A., Demel F., Schütz B., Rosner M., Kovacic B., Sukhbaatar N., Niederreiter B., Blüml S., Kuess P., Sexl V., Müller M., Mikula M., Weckwerth W., Haschemi A., Susani M., Hengstschläger M., Gambello M.J, & Weichhart T. (2017). Chronic signaling via the metabolic checkpoint kinase mTORC1 induces macrophage granuloma formation and marks sarcoidosis progression. Nature immunology, 18(3), 293-302.
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