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Vectastain elite dab kit

Manufactured by Vector Laboratories
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The Vectastain Elite DAB kit is a colorimetric detection system for use in immunohistochemistry and other immunodetection applications. It utilizes the 3,3'-diaminobenzidine (DAB) chromogen to produce a brown reaction product at the site of the target antigen.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (4 mentions) Horseradish peroxidase, by Thermo Fisher Scientific (3 mentions) Hrp conjugated secondary antibody, by Vector Laboratories (3 mentions) Anti p stat3, by Cell Signaling Technology (2 mentions) Anti mig 6, by Cell Signaling Technology (2 mentions) Anti ki67, by Abcam (2 mentions) Anti stat3, by Cell Signaling Technology (2 mentions) Biotinylated secondary antibody, by Vector Laboratories (2 mentions) Hematoxylin, by Biocare Medical (2 mentions) Proliferating cell nuclear antigen (pcna), by Agilent Technologies (1 mentions) Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (1 mentions) P jnk, by Agilent Technologies (1 mentions) P jnk, by Santa Cruz Biotechnology (1 mentions) Horseradish peroxidase conjugated secondary antibody, by Vector Laboratories (1 mentions) Normal goat serum (ngs), by Vector Laboratories (1 mentions) Anti ki67 ab15580, by Abcam (1 mentions) Anti e cadherin, by BD (1 mentions) Sc 6260, by Santa Cruz Biotechnology (1 mentions) Ab36495, by Abcam (1 mentions) Cd4 sc 1140, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

DAB kit (218 citations) Vectastain Elite kit (104 citations) Vectastain kit (77 citations) Vectastain Elite (56 citations) Vectastain DAB kit (12 citations) Vectastain Elite ABC and DAB Substrate kits (2 citations)

24 protocols using vectastain elite dab kit

1

Histological and Immunohistochemical Analysis

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Primary tumors, hearts, livers, lungs and kidneys were fixed overnight, embedded in paraffin and then cut into serial sections (4-μm thick). The sections were subjected to hematoxylin and eosin (H&E) staining. Representative images were acquired using a microscope (Leica). For immunohistochemical staining, slides were deparaffinized in xylene and rehydrated with graded alcohol and incubated in 3% H2O2 to block endogenous peroxidase activity. The slides were boiled for 30 min in 10 mM sodium citrate (pH 6.0) for antigen retrieval and then blocked in 5% BSA for 30 min, followed by incubation with antibodies against p-eif2α and ERO1-Lα at the appropriate dilution at 4 °C overnight in a humidity chamber. Afterward, the slides were washed three times with PBS and then incubated with secondary antibodies for 30 min at room temperature. Immunoreactivity was visualized using a Vectastain Elite DAB Kit (Vector Laboratories, Burlingame, CA, USA).
Wang Z., Yin F., Xu J., Zhang T., Wang G., Mao M., Wang Z., Sun W., Han J., Yang M., Jiang Y., Hua Y, & Cai Z. (2019). CYT997(Lexibulin) induces apoptosis and autophagy through the activation of mutually reinforced ER stress and ROS in osteosarcoma. Journal of Experimental & Clinical Cancer Research : CR, 38, 44.
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2

Immunohistochemical Analysis of Tumor Xenografts

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Paraffin-embedded tumor tissue of tumor xenografts was sectioned and immunohistochemically stained for PCNA, p-JNK, and cleaved caspase-3 using a kit from Dako (Copenhagen, Denmark). Following antibodies were bought from Santa Cruz: PCNA (sc-390003, 1:50, 4°C overnight), p-JNK (sc-390003, 1:50, 4°C overnight), and cleaved caspase-3 antibody (sc-390003, 1:50, 4 °C overnight). Vectastain Elite DAB KIT was bought from Vector Laboratories (CA, USA).
Li B., Zhou P., Xu K., Chen T., Jiao J., Wei H., Yang X., Xu W., Wan W, & Xiao J. (2020). Metformin induces cell cycle arrest, apoptosis and autophagy through ROS/JNK signaling pathway in human osteosarcoma. International Journal of Biological Sciences, 16(1), 74-84.
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3

Immunostaining Analysis of Uterine Tissues

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Immunostaining analyses were performed as previously described47 (link). Briefly, uterine sections were incubated
with appropriate primary antibodies, anti-MIG-6 (Customized antibody by Dr. Jeong Lab),
anti-pSTAT3 (CS-9131; Cell Signaling, Danvers, MA), anti-STAT3 (CS-4904; Cell Signaling,
Danvers, MA), and anti-Ki67 (ab15580; Abcam, Cambridge, MA), in 10% normal goat
serum in PBS overnight at 4°C. For immunohistochemistry, sections were incubated
with secondary antibody (Vector Laboratories, Burlingame, CA) and detected using the
Vectastain Elite DAB kit (Vector Laboratories, Burlingame, CA). For immunofluorescence,
sections were incubated with secondary antibody conjugated to Alexa Fluor 488-conjugated
anti-mouse IgG (Invitrogen Crop., Carlsbad, CA) for 2 hours at RT. Then, sections were
mounted with DAPI (Vector Laboratories, Burlingame, CA) to enable nuclear visualization.
The immunohistochemical staining intensities were graded by H-Score. The H-score was
calculated as previous reported48 (link).
Yoo J.Y., Yang W.S., Lee J.H., Kim B.G., Broaddus R.R., Lim J.M., Kim T.H, & Jeong J.W. (2017). MIG-6 negatively regulates STAT3 phosphorylation in uterine epithelial cells. Oncogene, 37(2), 255-262.
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4

Immunohistochemical Analysis of Tumor Samples

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Formalin-fixed tissue samples were embedded in paraffin and 4-μm sections were cut. Primary tumors, heart, liver, spleen, lung, and kidney sections were stained with H&E for routine histological examinations and morphometric analysis. For immunohistochemical staining, slides were deparaffinized in xylene and rehydrated with graded alcohol and incubated in 3% hydrogen peroxide to block the endogenous peroxidase activity. Antigen retrieval was performed by boiling the slides in 10 mM sodium citrate (pH 6.0) for 30 min. Then slides were blocked in 10% normal goat serum for 15 min, followed by incubation with p-ERK, and cleaved caspase-3 at 4 °C overnight in a moist chamber. On the next day, slides were washed in PBS and incubated with the second antibody for 1 h at room temperature. Immunoreactivity was detected using the Vectastain Elite DAB KIT (Vector Laboratories, Burlingame, CA, USA).
Huang K., Chen Y., Zhang R., Wu Y., Ma Y., Fang X, & Shen S. (2018). Honokiol induces apoptosis and autophagy via the ROS/ERK1/2 signaling pathway in human osteosarcoma cells in vitro and in vivo. Cell Death & Disease, 9(2), 157.
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5

Immunohistochemical Staining of Paraffin-Embedded Tissues

Cited 1 time
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Fixed, paraffin-embedded tissues were cut at 5 μm, mounted on slides, deparaffinized, and rehydrated in a graded alcohol series. For hematoxylin and eosin staining (H&E), slides were sequentially submerged in hematoxylin, 0.25% HCL, 1% lithium carbonate, and eosin, followed by dehydration and mounting. Immunostaining was performed as previously described (22 (link)) with specific commercially available primary antibodies (Table S1). For IHC, biotinylated secondary antibodies (Vector Laboratories, Burlingame, CA) were used, followed by incubation with horseradish peroxidase (Thermo Fisher Scientific, Waltham, MA) and developing using the Vectastain Elite DAB kit (Vector Laboratories). For immunofluorescence, appropriate species-specific fluorescently tagged secondary antibodies (Invitrogen, Carlsbad, CA) were used before mounting with DAPI (Vector Laboratories) for imaging. To compare the IHC staining intensities, a semiquantitative grade (H-score) was calculated by adding the percentage of strongly stained nuclei (3x), the percentage of moderately stained nuclei (2x), and the percentage of weakly stained nuclei (1x) in a region of approximately 100 cells, giving a possible range of 0–300. The percentage of Ki67-positive cells was counted in representative fields of approximately 150 epithelial cells and 150 stromal cells.
Marquardt R.M., Kim T.H., Yoo J.Y., Teasley H.E., Fazleabas A.T., Young S.L., Lessey B.A., Arora R, & Jeong J.W. (2020). Endometrial epithelial ARID1A is critical for uterine gland function in early pregnancy establishment. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 35(2), e21209.
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6

Immunohistochemical Analysis of Cell Proliferation

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The tissue specimens were fixed in 4% paraformaldehyde, then embedded in paraffin and sectioned. The tissue sections were deparaffinized in xylene, rehydrated with gradient alcohol, and boiled in sodium citrate for 30 min for antigen retrieval. Then, the tissue sections were blocked in 10% normal goat serum for 15 min and incubated with the primary antibodies against Ki-67 overnight at 4 °C. We washed the tissue sections in PBST and incubated them with the secondary antibody for 2 h at room temperature. A Vectastain Elite DAB kit (Vector Laboratories, Burlingame, CA, USA) was used to detect immunoreactivity.
Sun X., Zhao X., Xu S., Zhou Y., Jia Z, & Li Y. (2022). CircSRSF4 Enhances Proliferation, Invasion, and Migration to Promote the Progression of Osteosarcoma via Rac1. International Journal of Molecular Sciences, 23(11), 6200.
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7

Immunohistochemical Analysis of Formalin-Fixed Tissues

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Formalin-fixed tissue samples were embedded in paraffin and 4-μm sections were cut. Primary tumors, heart, liver, spleen, lung, and kidney sections were stained with H&E for routine histological examinations and morphometric analysis. For immunohistochemical staining, slides were deparaffinized in xylene and rehydrated with graded alcohol and incubated in 3% hydrogen peroxide to block the endogenous peroxidase activity. Antigen retrieval was performed by boiling the slides in 10 mM sodium citrate (pH 6.0) for 30 min. Then slides were blocked in 10% normal goat serum for 15 min, followed by incubation with PCNA, p-JNK, and cleaved caspase-3 at 4 °C overnight in a moist chamber. On the next day, slides were washed in PBS and incubated with the second antibody for 1 h at room temperature. Immunoreactivity was detected using the Vectastain Elite DAB KIT (Vector Laboratories, Burlingame, CA, USA).
Wang H., Zhang T., Sun W., Wang Z., Zuo D., Zhou Z., Li S., Xu J., Yin F., Hua Y, & Cai Z. (2016). Erianin induces G2/M-phase arrest, apoptosis, and autophagy via the ROS/JNK signaling pathway in human osteosarcoma cells in vitro and in vivo. Cell Death & Disease, 7(6), e2247-.
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8

Immunohistochemical Analysis of Arid1a in Uteri and Ovaries

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Three sections from each of three independent control and Arid1a cKO uteri or ovary were analyzed in parallel. Paraffin-embedded tissues were cut at 5 μm, mounted on silane-coated slides, deparaffinized, rehydrated in graded alcohol series, and boiled in citrate buffer, pH 6.0 (Abcam) prior to blocking with 10% normal goat serum and incubating in primary antibody diluted in 10% normal goat serum (Vector Laboratories) in phosphate-buffered saline (PBS), pH 7.5 (Life Technologies, Inc.) overnight at 4°C. Primary antibodies and conditions are listed in Supplemental Table S2. Sections were washed with PBS and incubated with the appropriate species-specific horseradish peroxidase-conjugated secondary antibody (2 μg/ml; Vector Laboratories) for 1 h at room temperature. Immunoreactivity was detected using the Vectastain Elite DAB kit (Vector Laboratories). Sections were then briefly counterstained with hematoxylin, dehydrated, and mounted. Slides were analyzed by light microscopy.
Wang X., Khatri S., Broaddus R., Wang Z, & Hawkins S.M. (2016). Deletion of Arid1a in Reproductive Tract Mesenchymal Cells Reduces Fertility in Female Mice. Biology of Reproduction, 94(4), 93.
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9

Immunohistochemical Analysis of Formalin-Fixed Tissues

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Formalin-fixed tissue samples were embedded in paraffin and 4 μm sections were cut. Primary tumors, heart, liver, spleen, lung, and kidney sections were stained with H&E for routine histological examinations and morphometric analysis. For immunohistochemical staining, slides were deparaffinized in xylene and rehydrated with graded alcohol and incubated in 3% hydrogen peroxide to block the endogenous peroxidase activity. Antigen retrieval was performed by boiling the slides in 10 mM sodium citrate (pH 6.0) for 30 min. Then, slides were blocked in 10% normal goat serum for 15 min, followed by incubation with p-JNK, and cleaved caspase-3 at 4 °C overnight in a moist chamber. On the next day, slides were washed in PBS and incubated with the second antibody for 1 h at room temperature. Immunoreactivity was detected using the Vectastain Elite DAB KIT (Vector Laboratories, Burlingame, CA, USA).
Shen S., Zhou M., Huang K., Wu Y., Ma Y., Wang J., Ma J, & Fan S. (2017). Blocking autophagy enhances the apoptotic effect of 18β-glycyrrhetinic acid on human sarcoma cells via endoplasmic reticulum stress and JNK activation. Cell Death & Disease, 8(9), e3055-.
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10

Immunohistochemical Analysis of ARG1 Expression

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Immunohistochemistry analyses were performed as previously described 40,41. Dewaxed hydrated paraffin-embedded tissue sections were blocked with 10% normal goat serum in PBS (pH 7.5) and then incubated with primary antibody diluted in 10% normal goat serum in PBS (pH 7.5) overnight at 4°C (anti-ARG1 (1:2000 dilution; ab32037; Abeam). On next day, the sections were incubated with the appropriate species specific HRP-conjugated secondary antibody (2 μg/ml; Vector Laboratories) for one hour at room temperature. Immunoreactivity was detected using the Vectastain Elite DAB kit (Vector Laboratories). A semiquantitative grading system (H-score) was used to compare the immunohistochemical staining intensities as previously described 42. The overall score ranged from 0 to 300.
Tran D.N., Rozen V., Hunter M.I., Kim T.H, & Jeong J.W. (2023). ARG1 is a potential prognostic marker in metastatic and recurrent endometrial cancer. Research Square.
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