Target retrieval solution s1699

Manufactured by Agilent Technologies

Sourced in Denmark

Target Retrieval Solution S1699 is a laboratory reagent used for antigen retrieval in immunohistochemistry (IHC) and immunocytochemistry (ICC) applications. It is designed to facilitate the exposure of target antigens in formalin-fixed, paraffin-embedded (FFPE) tissue sections or cell preparations, allowing for improved detection by primary antibodies.

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Lab products found in correlation

Immpress reagent, by Vector Laboratories (2 mentions) Normal rabbit serum, by Vector Laboratories (2 mentions) Glial fibrillary acidic protein (gfap), by Cell Signaling Technology (1 mentions) Ep1536y, by Abcam (1 mentions) Cd11b, by Abcam (1 mentions) Cd11b, by Agilent Technologies (1 mentions) Mhcii, by Agilent Technologies (1 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Southern Biotech (1 mentions) Bx60 microscope, by Olympus (1 mentions) Neuronal nuclei (neun), by Abcam (1 mentions) Ab211327, by Abcam (1 mentions) Ab32072, by Abcam (1 mentions) Bx41 microscope, by Olympus (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Neomarker rb 9013 p1, by Thermo Fisher Scientific (1 mentions) Dp12 camera, by Olympus (1 mentions) B 41 microscope, by Olympus (1 mentions) Immpress reagent anti rabbit ig peroxidase, by Vector Laboratories (1 mentions)

7 protocols using target retrieval solution s1699

Immunohistochemical Analysis of Mouse Brain

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Mice were euthanized by intra-cardiac perfusion and brains were fixed in neutral buffered formalin. Paraffin-embedded brain sections were assessed with the following antibodies: GFAP (Cell Signaling, 1:1000), Iba1 (Wako, 1:1000); pSer129/81A αSyn (1:3000; [13 (link), 36 (link)]); EP1536Y (AbCam, 1:1000); p62 (Protein Tech; 1:1000), cd11b (AbCam; 1:250), MHCII (Novus; 1:50) and NeuN (Abcam; 1:500). For all antibodies except cd11b and MHCII, antigen retrieval was performed by steaming for 25 min in water. For cd11b and MHCII, antigen retrieval was done by steaming in Dako Target Retrieval Solution S1699 (modified citrate buffer, pH 6.1). Colorimetric slides were treated with ImmPress reagents (Vector Labs) and visualized with 3, 3’diaminobenzidine followed by hematoxylin counterstaining. Fluorimetric slides were visualized with Alexa Fluor conjugated secondary antibodies (Invitrogen) and counterstaining with 4’, 6-diamidino-2-phenylindole (DAPI; Southern Biotech). All colorimetric slides were scanned using the Aperio XT whole slide scanner while fluorescent slides were visualized using Olympus BX60 microscope with a color digital camera.

Koller E.J., Brooks M.M., Golde T.E., Giasson B.I, & Chakrabarty P. (2017). Inflammatory pre-conditioning restricts the seeded induction of α-synuclein pathology in wild type mice. Molecular Neurodegeneration, 12, 1.

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Immunoperoxidase Analysis of Lung Tissue

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Lungs were perfused with 10% formalin, stored in fixative overnight, and embedded in paraffin. Four-micron thick sections of formalin fixed tissue were used for immunoperoxidase analysis after baking at 60 °C for 1 hour, deparaffinization and rehydration (100% xylene X4 for 3 minutes each, 100% ethanol X4 for 3 minutes each and running water for 5 minutes). The sections were blocked for peroxidase activity with 3% hydrogen peroxide in methanol for 10 minutes and washed under the running water for 5 minutes. The sections with pressure cooked (Biocare Medical) antigen retrieval were at 120°C i n Citrate Buffer (Dako Target Retrieval Solution, S1699). The slides were cooled for 15 minutes, and transferred to Tris buffer saline (TBS). The sections were incubated with rabbit monoclonal anti-MYC (Abcam Cat#ab32072; 1:900) and or rabbit monoclonal anti-ASCL1 antibody (Abcam Cat#ab211327; 1:100) was incubated 40 min room temperature. The secondary antibody was used Leica Novolink Polymer (Cat#RE7161) 30 min incubation. All the incubations were carried out in a humid chamber at room temperature. The slides were rinsed with TBS in between incubation. The sections were developed using 3,3’-diaminobenzidine (DAB) as substrate and counter-stained with Mayer’s Hematoxylin.

Zhang H., Christensen C.L., Dries R., Oser M.G., Deng J., Diskin B., Li F., Pan Y., Zhang X., Yin Y., Papadopoulos E., Pyon V., Thakurdin C., Kwiatkowski N., Jani K., Rabin A.R., Castro D.M., Chen T., Silver H., Huang Q., Bulatovic M., Dowling C.M., Sundberg B., Leggett A., Ranieri M., Han H., Li S., Yang A., Labbe K.E., Almonte C., Sviderskiy V.O., Quinn M., Donaghue J., Wang E.S., Zhang T., He Z., Velcheti V., Hammerman P.S., Freeman G.J., Bonneau R., Kaelin WG J.r., Sutherland K.D., Kersbergen A., Aguirre A.J., Yuan G.C., Rothenberg E., Miller G., Gray N.S, & Wong K.K. (2019). CDK7 Inhibition Potentiates Genome Instability Triggering Anti-Tumor Immunity in Small Cell Lung Cancer. Cancer cell, 37(1), 37-54.e9.

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Immunohistochemical Analysis of von Willebrand Factor

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For immunohistochemical analysis, the EnVision method was used according to Kloza et al. [55 (link)], using antibodies against von Willebrand factor (vWF) (1:2000, 2 h incubation at room temperature (RT), Polyclonal Rabbit Anti-Human (no cat. A 0082); DakoCytomation, Glostrup, Denmark). Antigen retrieval was performed before commencing immunohistochemical staining for vWF using Target Retrieval Solution (S1699; Dako, Glostrup, Denmark). A negative control was included, in which the antibody was replaced by normal rabbit serum (Vector Laboratories, Burlingame, CA, USA) at the respective dilution (no staining), and a positive control was included using rat lung stained for vWF. Immunohistochemical staining was evaluated on an Olympus BX41 microscope with an Olympus DP12 camera under 200× magnification.

Sadowska O., Baranowska-Kuczko M., Gromotowicz-Popławska A., Biernacki M., Kicman A., Malinowska B., Kasacka I., Krzyżewska A, & Kozłowska H. (2020). Cannabidiol Ameliorates Monocrotaline-Induced Pulmonary Hypertension in Rats. International Journal of Molecular Sciences, 21(19), 7077.

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IL-6 Expression in Mouse Brain

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Wild type B6/C3H mice received bilateral, intraventricular injections of rAAV-IL-6 on neonatal day P0 and were harvested at 1.5 months. Mice brains were divided sagitally; left hemisphere was snap frozen for biochemical determination of IL-6 levels and right hemisphere was fixed in formalin and paraffin embedded as coronal sections for IL-6 immunohistochemistry. The frozen hemibrains were extracted in RIPA buffer containing 1x Protease inhibitor cocktail (Roche, Indianapolis, IN) and used for biochemical determination of IL-6 protein levels using the Opti EIA mouse IL-6 assay (BD Biosciences) as described earlier [33 (link)]. Colorimetric assays were analyzed using the SoftMax data acquisition software (Molecular Devices). For immunohistochemistry, paraffin embedded slides were stained with anti IL-6 antibody (AbCam, 1:200) following antigen retrieval by steaming in Dako Target Retrieval Solution S1699 (modified citrate buffer, pH 6.1). Colorimetric slides were treated with ImmPress reagents (Vector Labs) and visualized with 3, 3’diaminobenzidine followed by hematoxylin counterstaining.

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Immunohistochemical Evaluation of Oral Biopsies

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Oral biopsies were fixed en bloc in 10% neutral buffered formalin, embedded in paraffin, sectioned, mounted, and stained with H&E according to standard laboratory protocols. Immunohistochemistry was performed on 4‐µm thick, formalin‐fixed, paraffin‐embedded tissue sections, as previously described, with antigen retrieval performed in Dako Target Retrieval Solution (S1699) for 30 minutes at 95°C 1. Primary antibodies and dilutions were: rat anti‐CD3 (clone 3–12, diluted 1:10, Leukocyte Antigen Biology Lab, UCD); and rabbit anti‐CD20 (NeoMarker RB‐9013‐P1; 1:300; Thermo Fisher Scientific, Pittsburgh). Sections were then incubated with a Streptavidin‐HRP label (anti‐rabbit link, or anti‐rat link; Biocare Medical's 4+ Detection System GR608, or GR607 and HP604, respectively). Detection was visualized with Vector NovaRED for peroxidase (SK‐4800), per manufacturer's instructions. Sections were counterstained in Mayer's hematoxylin. Nonspecific background was evaluated with duplicate sections that received diluent in place of the primary antibody. Biopsies were interpreted by a board‐certified veterinary pathologist (NV).

Arzi B., Clark K.C., Sundaram A., Spriet M., Verstraete F.J., Walker N.J., Loscar M.R., Fazel N., Murphy W.J., Vapniarsky N, & Borjesson D.L. (2017). Therapeutic Efficacy of Fresh, Allogeneic Mesenchymal Stem Cells for Severe Refractory Feline Chronic Gingivostomatitis. Stem Cells Translational Medicine, 6(8), 1710-1722.

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Immunohistochemistry for von Willebrand Factor

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In the immunohistochemical study, the EnVision method was used according to Tokajuk et al. [60 (link)] using antibodies against von Willebrand factor (vWF) (1:2000–2 h incubation at RT, Polyclonal Rabbit Anti-Human (no cat. A 0082); DakoCytomation, Glostrup, Denmark). Antigen retrieval was performed before commencing immunohistochemical staining for vWF using a Target Retrieval Solution (S1699; Dako, Denmark). A negative control was included in which the antibody was replaced by normal rabbit serum (Vector Laboratories, Burlingame, CA, USA) at the respective dilution (no staining), and a positive control was included using rat lung stained for vWF. The obtained results of immunohistochemical staining were evaluated on an Olympus B×41 microscope with an Olympus DP12 camera under 200× magnification. The intensity of the immunohistochemical reaction was measured using a 0 to 256 grey scale level in which completely black pixels were given a value of 256 and white or bright pixels were given a value of 0.

Kloza M., Baranowska-Kuczko M., Toczek M., Kusaczuk M., Sadowska O., Kasacka I, & Kozłowska H. (2019). Modulation of Cardiovascular Function in Primary Hypertension in Rat by SKA-31, an Activator of KCa2.x and KCa3.1 Channels. International Journal of Molecular Sciences, 20(17), 4118.

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Immunohistochemical Analysis of BRK Protein in Breast Cancer

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Immunohistochemistry (IHC) staining on human breast cancer tissue array BR10010a and BR243d with rabbit anti-BRK (C-18) antibody was performed and analyzed by USBIOMAX (https://www.biomax.us/). Briefly, the tissue samples on each array were formalin fixed, paraffin embedded. Tissue array sections were mounted on the positive charged SuperFrost Plus glass slide. Primary antibody rabbit anti-BRK(C-18) antibody (sc-1188) was purchased from Santa Cruz Biotechnology, Inc. ImmPRESS™ Reagent anti-Rabbit Ig (peroxidase) of catalog number MP7401 were purchased from Vector Laboratories. DAB (DAKO Cytomation, Code K3465) used as substrate chromogen. Antigen retrieval solution was purchased from DakoCytomation (Target Retrieval solution, S-1699). The standard procedure can be obtained through https://www.biomax.us/.

Miah S., Bagu E., Goel R., Ogunbolude Y., Dai C., Ward A., Vizeacoumar F.S., Davies G., Vizeacoumar F.J., Anderson D, & Lukong K.E. (2019). Estrogen receptor signaling regulates the expression of the breast tumor kinase in breast cancer cells. BMC Cancer, 19, 78.

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