Ar9 buffer

Manufactured by Akoya Biosciences

The AR9 buffer is a specialized laboratory reagent developed by Akoya Biosciences. It is designed to facilitate specific experimental protocols. The core function of the AR9 buffer is to maintain the stability and integrity of samples during various analytical and processing steps. No further details or interpretations are provided.

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Lab products found in correlation

Decloaker, by Biocare Medical (2 mentions) Bond polymer refine detection kit, by Leica (2 mentions) Rat anti mouse f4 80, by Bio-Rad (1 mentions) Opal 520 reagent pack, by PerkinElmer (1 mentions) Opal 570 reagent pack, by PerkinElmer (1 mentions) Primary rat antimouse f4 80 antibody, by Bio-Rad (1 mentions) Rabbit antimouse ym1, by STEMCELL (1 mentions) Blocking diluent buffer, by PerkinElmer (1 mentions) Eclipse 90i, by Nikon (1 mentions) Sp8 lightning confocal microscope, by Leica (1 mentions) Cd11b, by Abcam (1 mentions) Cd11b, by BD (1 mentions) Rabbit anti pd l1, by Cell Signaling Technology (1 mentions) Rabbit anti pd 1, by Cell Signaling Technology (1 mentions) Rabbit anti cd11b, by Cell Signaling Technology (1 mentions) Opal polymer hrp ms rb, by PerkinElmer (1 mentions) Opal 7 color manual ihc kit, by PerkinElmer (1 mentions) Ar6 buffer, by Akoya Biosciences (1 mentions) F6057, by Merck Group (1 mentions)

Close spelling variants of this product

AR9 retrieval buffer

5 protocols using ar9 buffer

Histopathological Analysis of Immune Infiltrates

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Histopathological services were provided by UCLA Translational Pathology Core Lab. Mice footpad samples were processed, decalcified and sectioned for H&E staining, and subsequent image analysis. Immunohistochemistry stainings were also performed on these footpad tissues: Paraffin-embedded sections were cut at 4-μm thickness and paraffin was removed with xylene and the sections were rehydrated through graded ethanol. Endogenous peroxidase activity was blocked with 3% hydrogen peroxide in methanol for 10 min. Heat-induced antigen retrieval was carried out for all sections in AR9 buffer (AR9001KT Akoya) using a Biocare decloaker at 95°C for 25 min. The slides were then stained with primary antibodies targeting mouse F4/80, CD4 and CD8 antigens at 4°C overnight; the signal was detected using Bond Polymer Refine Detection Kit (Leica Microsystems, catalog #DS9800) with a diaminobenzidine reaction to detect antibody labeling and hematoxylin counterstaining.

Garcia G J.r., Irudayam J.I., Jeyachandran A.V., Dubey S., Chang C., Castillo Cario S., Price N., Arumugam S., Marquez A.L., Shah A., Fanaei A., Chakravarty N., Joshi S., Sinha S., French S.W., Parcells M.S., Ramaiah A, & Arumugaswami V. (2023). Innate immune pathway modulator screen identifies STING pathway activation as a strategy to inhibit multiple families of arbo and respiratory viruses. Cell Reports Medicine, 4(5), 101024.

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Immunohistochemical Analysis of Murine Macrophages

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Formalin-fixed, paraffin-embedded tissue sections (pancreatic body and tail; female and male mice combined) were dewaxed with xylene and rehydrated in an ethanol series. Antigen retrieval was performed in AR9 buffer (Akoya Biosciences, Menlo Park, CA), and sections were then immersed in blocking/diluent buffer (PerkinElmer, Richmond, CA). For immunohistochemistry, sections were incubated overnight with primary rat antimouse F4/80 antibody (#MCA497B; Bio-Rad) diluted 1:200 in blocking/diluent buffer. After washing, the Labelled Polymer-Dako REAL EnVision HRP kit (Agilent Dako, Santa Clara, CA) was then used to visualize antigen-antibody complexes.
For immunofluorescence, sections were incubated overnight with rat antimouse F4/80 (#MCA497B; Bio-Rad), rabbit antimouse pSTAT (Tyr701) (#44-376; Thermo Fisher Scientific), and rabbit antimouse Ym1 (#60130; STEMCELL Technologies, Cambridge, MA) antibodies (1:200 dilution in blocking buffer). Slides were then counterstained with multiplexing fluorescence kits from PerkinElmer: Opal 520 Reagent Pack, FITC, Opal 570 Reagent Pack, TRITC, and Opal 650 Reagent Pack, CY5, according to manufacturer’s instructions. 4′,6-diamidino-2-phenylindole was used for nuclear counterstain. Images were taken with a Nikon Eclipse 90i.

Ako S., Teper Y., Ye L., Sinnett-Smith J., Hines O.J., Rozengurt E, & Eibl G. (2022). Statins Inhibit Inflammatory Cytokine Production by Macrophages and Acinar-to-Ductal Metaplasia of Pancreatic Cells. Gastro hep advances, 1(4), 640-651.

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Histopathological Analysis of Immune Cells in Mouse Footpad

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Histopathological services were provided by UCLA Translational Pathology Core Lab. Mice footpad samples were processed, decalcified and sectioned for H&E staining, and subsequent image analysis. Immunohistochemistry stainings were also performed on these footpad tissues: Paraffin-embedded sections were cut at 4-μm thickness and paraffin was removed with xylene and the sections were rehydrated through graded ethanol. Endogenous peroxidase activity was blocked with 3% hydrogen peroxide in methanol for 10 minutes. Heat-induced antigen retrieval was carried out for all sections in AR9 buffer (AR9001KT Akoya) using a Biocare decloaker at 95°C for 25 minutes. The slides were then stained with primary antibodies targeting mouse F4/80, CD4 and CD8 antigens at 4°C overnight; the signal was detected using Bond Polymer Refine Detection Kit (Leica Microsystems, catalogue #DS9800) with a diaminobenzidine reaction to detect antibody labeling and hematoxylin counterstaining.

Garcia G J.r., Irudayam J.I., Jeyachandran A.V., Dubey S., Chang C., Cario S.C., Price N., Arumugam S., Marquez A.L., Shah A., Fanaei A., Chakravarty N., Joshi S., Sinha S., French S.W., Parcells M., Ramaiah A, & Arumugaswami V. (2023). Broad-spectrum antiviral inhibitors targeting pandemic potential RNA viruses. bioRxiv.

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Multicolor IF Staining of Mouse Colon

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Immunofluorescence staining of mouse colons was performed by Applied Pathology Systems, LLC. Formalin-Fixed Paraffin-Embedded (FFPE) colon tissues were sectioned and mounted to glass slides. Tissue sections were dewaxed, rehydrated, and subjected to the antigen retrieval in microwave with AR9 Buffer (Akoya Biosciences). Slides were then blocked with Blox All blocking buffer (Vector Laboratories) and Opal antibody block reagent (Akoya Biosciences) prior to the co-staining. The co-staining of STING (CST) with CD3 (Genetex) or CD11b (Abcam) was performed using the Opal 4 color manual IHC kit (NEL810001KT). STING was labeled with Opal 690 reagent while CD3 or CD11b was labeled with Opal 480 reagent FITC conjugated E-Cadherin antibody (BD Biosciences) was used in the STING and E-Cadherin co-staining. Fluorescence co-stained slides were cover slipped with VectorSheild Mounting media (Vector Laboratories) before imaging with Leica SP8 Lightning Confocal Microscope.

Shmuel-Galia L., Humphries F., Lei X., Ceglia S., Wilson R., Jiang Z., Ketelut-Carneiro N., Foley S.E., Pechhold S., Houghton J., Muneeruddin K., Shaffer S.A., McCormick B.A., Reboldi A., Ward D., Marshak-Rothstein A, & Fitzgerald K.A. (2021). Dysbiosis exacerbates colitis by promoting ubiquitination and accumulation of the innate immune adaptor STING in myeloid cells. Immunity, 54(6), 1137-1153.e8.

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Multiplex Immunofluorescence Protocol for DLBCL TMA Analysis

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The DLBCL TMA slides were stained with multiplex uorescence by using the Opal 7-color Manual IHC Kit (PerkinElmer, MA). After dewaxing by xylene and rehydration by ethanol, slides were heated in a microwave with AR6 Buffer (AR600, AKOYA) and AR9 Buffer (AR900, AKOYA) for antigen retrieval. The slides were incubated with primary antibodies overnight at 4°C and then incubated with secondary antibody for 10min at room temperature. At last, we used 4',6-diamidino-2-phenylindole (DAPI; F6057, Sigma) to stain the nuclei and seal the slides. Imaging was achieved using the Vectra 3.0 Automated Quantitative Pathology Imaging System. Tumor and stroma images were captured at ×20 magni cation. Finally, the staining was scored by inForm® Cell Analysis software based on the intensity and degree of staining.
The primary antibodies used in this study were as follows: rabbit anti-PHKA1 (24279-1-AP, Proteintech), rabbit anti-PLTP (ab282456, Abcam), rabbit anti-CD163 (93498, Cell Signaling Technology), rabbit anti-CD68 (76437, Cell Signaling Technology), rabbit anti-CD11B (49420, Cell Signaling Technology), mouse anti-CD66b (ARG66287, Arigobio), rabbit anti-PD-1 (86163, Cell Signaling Technology) and rabbit anti-PD-L1 (13684, Cell Signaling Technology). The secondary antibody was Opal™ polymer HRP Ms+Rb (ARH1001EA, Perkin Elmer).

He J., Chen Z., Xue Q., Sun P., Wang Y., Zhu C., & Shi W. (2022). Identification of molecular subtypes and a novel prognostic model of diffuse large B-cell lymphoma based on a metabolism-associated gene signature.

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