Retrievagen a

Manufactured by BD

Sourced in United States

Retrievagen A is a laboratory equipment product. It is designed to assist in the retrieval and extraction of biological samples from various sources. The core function of Retrievagen A is to facilitate the efficient and reliable recovery of materials for further analysis or processing.

Automatically generated - may contain errors

Lab products found in correlation

Rabbit anti grp78, by Santa Cruz Biotechnology (1 mentions) Vectastain avidin biotin complex kit, by Vector Laboratories (1 mentions) Biotinylated anti brdu antibody, by BD (1 mentions) B5002, by Merck Group (1 mentions) Brdu in situ detection kit, by BD (1 mentions) Envision dual link system hrp, by Agilent Technologies (1 mentions) G9023, by Merck Group (1 mentions) Clone g168 15, by BD (1 mentions) Anti brdu antibody, by BD (1 mentions) Biozero bz x710, by Keyence (1 mentions) Vectashield hardset mounting medium with dapi, by Vector Laboratories (1 mentions) Anti cd68, by Wuhan Servicebio Technology (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Wuhan Servicebio Technology (1 mentions) Mounting medium, by Thermo Fisher Scientific (1 mentions) Novared, by Vector Laboratories (1 mentions) Eclipse 80i microscope, by Nikon (1 mentions) Nis elements software, by Nikon (1 mentions) Lsm 710 confocal scanning microscope, by Zeiss (1 mentions) Autostainer xl, by Nikon (1 mentions) Inverted microscope, by Nikon (1 mentions)

15 protocols using retrievagen a

Endometrial GRP78 Expression Evaluation

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Archival endometrial samples were evaluated for GRP78 as previously described [11 (link),12 (link)]. Briefly, freshly cut formalin-fixed, paraffin-embedded samples were deparaffinized and rehydrated. Antigen retrieval was accomplished with BD Retrievagen A (BD Pharmingen, Carpenteria, CA) according to manufacturer instructions. GRP78 was detected with rabbit anti-GRP78 (1:100, Santa Cruz Biotechnologies, Santa Cruz, CA) followed by the goat anti-rabbit antibody (VECTASTAIN® avidin-biotin complex (ABC) kit, Vector Laboratories, Burlingame, CA). Visualization was achieved with 3,3′-diaminobenzidine (DAB).
The study pathologist (PMF), blinded to clinical and treatment data, scored all specimens as negative, weak, moderate and strong. Negative controls absent of primary antibody were used as a reference. Overexpression of GRP78 was defined as moderate/strong intensity [12 (link)]. Internal consistency was evaluated by re-presenting a random subset of previously scored slides to the same pathologist, blinded to the original score.

Tierney K.E., Ji L., Dralla S.S., Yoo E., Yessaian A., Pham H.Q., Roman L., Sposto R., Mhawech-Fauceglia P, & Lin Y.G. (2016). Endoplasmic reticulum stress in complex atypical hyperplasia as a possible predictor of occult carcinoma and progestin response. Gynecologic oncology, 143(3), 650-654.

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BrdU Cell Proliferation Assay in Liver

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BrdU incorporation assay was done using BrdU In-Situ Detection Kit (550803, BD Pharmingen) according to the manufacturer’s instructions. In brief, 2 hr prior to euthanasia mice were injected intraperitoneally with BrdU (B5002, Sigma Aldrich), at a dose of 50 mg/kg body weight of mice. Tissue samples were fixed in 4% paraformaldehyde and embedded in paraffin. The paraffin-embedded liver tissue sections were de-paraffinized by washing with xylene 2 times for 5 min each time at room temperature. The tissue sections were dehydrated by incubation in 100% ethanol 2 times for 5 min followed by once in 95% ethanol for 3 min at room temperature. The tissue sections were treated with 0.3% H₂O₂ to block endogenous peroxidase, followed by antigen retrieval with ‘BD Retrievagen A’ (#550803, BD Pharmingen) in a microwave oven to 89°C for 10 min. The tissue sections were incubated in biotinylated anti-BrdU antibody (#550803, BD Pharmingen) at 1:10 in diluent buffer for 1 hr at room temperature, and then in HRP-conjugated streptavidin for 1 hr at room temperature. The tissue sections were stained with DAB substrate solution followed by hematoxylin counterstaining. Slides were examined by bright field microscopy.

Udden S.N., Kwak Y.T., Godfrey V., Khan M.A., Khan S., Loof N., Peng L., Zhu H, & Zaki H. (2019). NLRP12 suppresses hepatocellular carcinoma via downregulation of cJun N-terminal kinase activation in the hepatocyte. eLife, 8, e40396.

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Immunohistochemical Analysis of MLH1 and MSH2

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Sections were dewaxed in xylene and rehydrated through a graded series of ethanol concentrations (100, 95 and 70%) to water. Antigen retrieval was performed by immersing the slides in BD Retrievagen A (pH 6.5 for MLH1 and pH 6.0 for MSH2; BD Biosciences, San Jose, CA, USA) and heating the slides in a microwave oven for 30 min at 95°C. Sections were treated with 3% hydrogen peroxide for 5 min to block endogenous peroxidase activity. Sections were washed in PBS and subsequently placed in 20% normal goat serum (G9023; Sigma-Aldrich; Merck Millipore) in PBS for 20 min to reduce non-specific staining. Sections were then incubated with monoclonal antibodies raised against human (h) MLH1 (1:10; clone G168-15; BD Biosciences) for 1 h 32 min at 42°C or against MSH2 (1:25; clone 25D12; Thermo Fisher Scientific, Inc.) for 1 h at room temperature. Subsequently, visualization was performed using the EnVision+Dual Link system-HRP (Dako) according to the manufacturer's protocol, using 3,3′-diaminobenzidine as a chromogen. Finally, slides were counterstained with hematoxylin solution. Both MLH1 and MSH2 were scored as either negative or positive staining. Tissue specimens were analyzed by two independent pathologists blinded to the conditions.

Huang C.J., Huang S.H., Chien C.C., Lee H.H., Yang S.H., Chang C.C, & Lee C.L. (2016). Impact of microsatellite status on chemotherapy for colorectal cancer patients with KRAS or BRAF mutation. Oncology Letters, 12(6), 4427-4434.

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BrdU Proliferation Assay in Mice

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Mice were injected intraperitoneally with bromodeoxyuridine (BrdU; BD Pharmingen) (100 mg/kg) 2.5 h prior to euthanasia. The intestine was isolated, formalin fixed, jelly-rolled, and paraffin embedded. Tissue sections were deparaffinized using BD^™ Retrievagen A and stained with anti-BrdU antibody according to the manufacturer’s instructions (BD Biosciences).

Keerthivasan S., Aghajani K., Dose M., Molinero L., Khan M.W., Venkatesvaran V., Weber C., Emmanuel A.O., Sun T., Ramos E.M., Keshavarzian A., Mulcahy M., Blatner N., Khazaie K, & Gounari F. (2014). Wnt/β-catenin signaling in T-cells drives epigenetic imprinting of pro-inflammatory properties and promotes colitis and colon cancer. Science translational medicine, 6(225), 225ra28.

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Immunofluorescence Staining of E-cadherin

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Tissue sections above mentioned were deparaffinized with xylene, and rehydrated. This was followed by heat-induced antigen retrieval in BD Retrievagen A (pH 6.0, BD Biosciences, San Diego CA, USA). After blocking in fetal bovine serum (FBS), tissues were incubated with fluorescence-labeled anti-E-cadherin antibody (Alexa FluorTM 594 anti-mouse/human CD324, Biolegend, San Diego, CA, USA). After mounting with VECTASHIELD HardSet Mounting Medium with DAPI (Vector Laboratories, ABurlingame CA, USA), the stained sections were visualized by fluorescence microscopy (Biozero BZ-X710; Keyence Corporation, Osaka, Japan).

Yamazaki K., Kato T., Tsuboi Y., Miyauchi E., Suda W., Sato K., Nakajima M., Yokoji-Takeuchi M., Yamada-Hara M., Tsuzuno T., Matsugishi A., Takahashi N., Tabeta K., Miura N., Okuda S., Kikuchi J., Ohno H, & Yamazaki K. (2021). Oral Pathobiont-Induced Changes in Gut Microbiota Aggravate the Pathology of Nonalcoholic Fatty Liver Disease in Mice. Frontiers in Immunology, 12, 766170.

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Colon Tissue Analysis in Mice

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Colon tissues of mice were fixed in Methanol–Carnoy’s fixative without washing, embedded in paraffin, and sectioned. Sections were dewaxed, hydrated, retrieved by RetrievagenA (BD Biosciences, USA). For IHC, sections were stained with anti-CD68 (Servicebio, China) antibodies. For IF, sections were stained with FITC-anti-MUC2, Cy3-anti-MPO, FITC-anti-CD4, Cy3-anti-CD8 (Abcam, UK) antibodies and DAPI (Servicebio, China). PAS staining was performed to visualize goblet cells and mucus layer in sections. Pictures were obtained by fluorescence microscope (Nikon Eclipse TI-SR, Japan) and CaseViewer2.4system (3DHISTECH, Hungary).

Xie B., Wang B., Shang R., Wang L., Huang X, & Xie L. (2023). Blocking MyD88 signaling with MyD88 inhibitor prevents colitis-associated colorectal cancer development by maintaining colonic microbiota homeostasis. Scientific Reports, 13, 22552.

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Immunohistochemical Quantification of Neutrophils

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Formalin-fixed paraffin sections of small intestine were subjected to rehydration, and endogenous peroxidase activity was quenched with 3% H₂O₂. Then antigen retrieval was performed using Retrievagen A (BD Pharmingen, San Jose, CA) according to the manufacturer’s directions. The sections were blocked with 10% BSA/PBS containing the serum from host species of secondary antibody. The primary antibody rat-anti-mouse Ly-6B.2 (Gr1) clone 7/4 at 1/250 dilution (Bio-Rad, Herculese, CA) was used to stain for neutrophils. Primary antibody or isotype control antibody prepared in 10% BSA/PBS were applied overnight at 4°C. The slides were then incubated with horseradish peroxidase (HRP)-conjugated secondary antibody for 60 min at room temperature, developed with NovaRED (Vector Laboratories, Burlingame, CA), counterstained with hematoxylin, and dehydrated. The sections were mounted in mounting medium (Thermo Scientific, Waltham, MA) and evaluated with Nikon eclipse 80i microscope. Images were analyzed using Nikon NIS-Elements software (Nikon, Melville, NY). For neutrophil infiltration positive staining cells in the area of injury were counted in 20 high power fields (HPF) at 200× and the average was calculated and expressed as number of neutrophils per HPF.

Geha M., Tsokos M.G., Bosse R.E., Sannikova T., Iwakura Y., Dalle Lucca J.J., De Waal Malefyt R, & Tsokos G.C. (2017). IL-17A produced by innate lymphoid cells is essential for intestinal ischemia reperfusion injury: ILC derived IL-17A is essential for intestinal IR injury. Journal of immunology (Baltimore, Md. : 1950), 199(8), 2921-2929.

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Histological Characterization of Engineered Constructs

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On day 28, constructs (n=2) were fixed in 10% formalin for 30 min at RT, then snap frozen and cryosectioned. Sections were stained for safranin-O or masson's trichrome on a Leica autostainer XL and imaged in bright field (40X objective) on a Nikon inverted microscope.
For immunostaining, sections were blocked with 10% goat serum, then analyzed by anti-collagen type II (1:50, US Biologicals) and anti-collagen type I (1:50). Sections were treated with appropriate enzymes for 1 hour at 37 °C: hyaluronidase (2080 U) for collagen II, and pepsin A (4000 U) with Retrievagen A (BD Biosciences) treatment for collagen I to help expose the antigen. Sections were probed with AlexaFluor 568-conjugated secondary antibodies and counterstained with DAPI for cell nuclei. All samples were processed at the same time to minimize sample-to-sample variation. Images were collected on a Zeiss LSM710 scanning confocal microscope with a 20X objective using the same settings and post-processing for all images. The background gain was set to negative controls on blank sections that received the same treatment. Positive controls were performed on porcine hyaline cartilage for collagen type II and porcine meniscus for collagen type I (Supplemental Figure 1).

Sridhar B.V., Doyle N.R., Randolph M.A, & Anseth K.S. (2014). Covalently tethered TGF-β1 with encapsulated chondrocytes in a PEG hydrogel system enhances extracellular matrix production. Journal of biomedical materials research. Part A, 102(12), 4464-4472.

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Immunohistochemical Detection of EPS8 in Mouse Tumors

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Detection of EPS8 in mouse tumour sections was carried out as follows. Five-micrometre tissue sections were dewaxed in Safe-Clear (Fisher Scientific, Pittsburgh, PA) and rehydrated, and antigen retrieval was performed by incubation in Retrievagen A (BD Biosciences, CA) at 95 °C for 30 min. After cooling to ambient temperature, endogenous peroxidase was blocked by incubation in 3% hydrogen peroxide for 15 min; slides were washed in TBS pH 8.0 and then blocked using a commercially available kit (M.O.M. Immunodetection Kit—Peroxidase; Vector Laboratories, Burlingame, CA). Sections were then incubated with EPS8 antibody (12.5 μg/ml) or the equivalent concentration of normal mouse IgG as control at ambient temperature for 1 h, washed in TBS and then incubated sequentially with biotinylated secondary antibody and streptavidin peroxidase reagent, developed using 3, 3’-diaminobenzidine substrate, counterstained with haematoxylin, dehydrated, mounted and imaged by brightfield microscopy (Keyence Corporation, Itasca, IL).

Shahoumi L.A., Khodadadi H., Bensreti H., Baban B, & Yeudall W.A. (2020). EPS8 phosphorylation by Src modulates its oncogenic functions. British Journal of Cancer, 123(7), 1078-1088.

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Immunohistochemical Staining of CD45+ Cells

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Tissue sections (3 μm) were cut from 4% paraformaldehyde (PFA)-fixed paraffin-embedded recipient organs. Sections were deparaffinized using xylene and ethanol and antigen retrieval was performed (Retrievagen A (pH 6.0), BD Pharmingen^TM). Non-specific background was reduced by incubating the slides in methanol + H₂O₂ (Wako). After blocking with horse serum, slides were incubated with mouse anti-human CD45 antibody (DAKO, M0701) (1:150) then HRP-conjugated horse anti-rabbit/mouse IgG antibody (ImmPRESS^TM, Cat. MP-7500). Slides were then stained with 3,3’-diaminobenzine (DAB) and hematoxylin, dehydrated, mounted using Vectamount (Vector Laboratories) and analyzed with Axiovert 200 microscope (Zeiss). Photos were taken using AxioCam MRc 5 (Zeiss) using the AxioVision rel. 4.6 software.

de Groot A.P., Saito Y., Kawakami E., Hashimoto M., Aoki Y., Ono R., Ogahara I., Fujiki S., Kaneko A., Sato K., Kajita H., Watanabe T., Takagi M., Tomizawa D., Koh K., Eguchi M., Ishii E., Ohara O., Shultz L.D., Mizutani S, & Ishikawa F. (2021). Targeting critical kinases and anti-apoptotic molecules overcomes steroid resistance in MLL-rearranged leukaemia. EBioMedicine, 64, 103235.

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