Mouse anti rpe65

Manufactured by Novus Biologicals

Sourced in United States

Mouse anti-RPE65 is an antibody that recognizes the RPE65 protein. RPE65 is a retinal pigment epithelium-specific protein that plays a key role in the visual cycle. This antibody can be used for the detection and analysis of RPE65 in various applications such as western blotting, immunohistochemistry, and flow cytometry.

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Lab products found in correlation

Goat anti pcadherin, by R&D Systems (1 mentions) Horseradish peroxidase hrp conjugated goat anti mouse and goat anti rabbit antibodies, by Thermo Fisher Scientific (1 mentions) Mouse anti β actin, by Merck Group (1 mentions) Rabbit anti gapdh, by Cell Signaling Technology (1 mentions) Alexa fluor 488 labeled goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Rabbit anti zo 1, by Santa Cruz Biotechnology (1 mentions) Mouse anti na k atpase, by Santa Cruz Biotechnology (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Ab5076, by Abcam (1 mentions) Nb100 355, by Novus Biologicals (1 mentions) Goat anti iba1, by Abcam (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488, by Jackson ImmunoResearch (1 mentions) Hrp conjugated donkey anti mouse igg, by Jackson ImmunoResearch (1 mentions) Anti sod1, by Proteintech (1 mentions) Hrp conjugated donkey anti rat igg, by Jackson ImmunoResearch (1 mentions) Hrp conjugated donkey anti rabbit igg, by Jackson ImmunoResearch (1 mentions) Rabbit anti calreticulin, by Cell Signaling Technology (1 mentions) Anti catalase, by Proteintech (1 mentions) Rabbit anti syntenin 1, by Abcam (1 mentions)

4 protocols using mouse anti rpe65

Antibody Detection in Tissue Samples

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Primary antibodies used were: mouse anti-β-Actin (A2228; Sigma-Aldrich, St. Louis, MO, USA), goat anti-p-cadherin (AF761, R&D Systems, Minneapolis, MN, USA), rabbit anti-MCT3 [140 (link)], rabbit-anti-GAPDH (#5174, Cell Signaling Technology, Danvers, MA, USA), and mouse anti-RPE65 (NB100-355, Novus Biologicals LLC, Centennial, CO, USA). Secondary antibodies used were: goat anti-mouse and goat anti-rabbit horseradish peroxidase (HRP)-conjugated antibodies (Thermo Fisher Scientific, Waltham, MA, USA), donkey anti-goat, and anti-rabbit IgG Alexa Fluor 594/488 conjugates (Invitrogen, Waltham, MA, USA).

Dhingra A., Tobias J.W., Philp N.J, & Boesze-Battaglia K. (2023). Transcriptomic Changes Predict Metabolic Alterations in LC3 Associated Phagocytosis in Aged Mice. International Journal of Molecular Sciences, 24(7), 6716.

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Immunocytochemical Analysis of ARPE-19 Cells

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After fixed with 4% paraformaldehyde for 15 min at room temperature, ARPE-19 cells were washed with PBS containing 0.3% Triton X-100 (PBST) three times and blocked at 37 °C for 1 h in PBST supplemented with 5% FBS (Gibco). Then, the cells were incubated with the following primary antibodies at 4 °C overnight: rabbit anti-ZO-1 (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), mouse anti-Na⁺K⁺ATPase (Santa Cruz Biotechnology Inc.), and mouse anti-RPE-65 (Novus Biologicals, Littleton, CO, USA). Cells were washed 3 times for 5 min with PBST and incubated with Alexa Fluor 488-labeled goat anti-rabbit IgG (Molecular Probes, Eugene, OR, USA) and Alexa Fluor 635-labeled goat anti-mouse IgG (Molecular Probes) for 1 h at room temperature. Stained cells were examined under Olympus FV1000 Confocal Scanning Scope (Olympus, Tokyo, Japan).

Song J., Han D., Lee H., Kim D.J., Cho J.Y., Park J.H, & Seok S.H. (2020). A Comprehensive Proteomic and Phosphoproteomic Analysis of Retinal Pigment Epithelium Reveals Multiple Pathway Alterations in Response to the Inflammatory Stimuli. International Journal of Molecular Sciences, 21(9), 3037.

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Choroid Immunohistochemistry Protocol

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After enucleating the eyes, the anterior segments were removed and the retina was carefully excised. Eyecups were then fixed overnight with 2% PFA at 4°C. After washing, four pie cuts were made to allow flattening of the choroid/sclera eyecup as well as clearly isolate superior and inferior quadrants. Then the choroid was prepared for IHC as previously described (19 (link)). The choroid was blocked with 2% normal goat or donkey serum for 4 h at 4°C, washed in 0.1% triton X-100 in TBS, and then incubated with a mixture of the following primary Abs: mouse anti-RPE65 (1:200, NB100–355, Novus Biologicals, Centennial, CO, USA), goat anti-Iba1 (1:200, ab5076, Abcam, Cambridge, MA, USA) and rabbit anti-MC tryptase (1:200, CAU26568, Biomatik, Wilmington, DE, USA) overnight at 4°C. After washing, they were incubated with Alexa-Fluor 488 or Cy3 conjugated goat or donkey secondary Abs (1:300, Jackson ImmunoResearch, West Grove, PA, USA) overnight, followed by mouse anti-ZO-1 Ab conjugated with Alexa-Fluor 594 (1:100, 339194, Thermo Fisher Scientific, Waltham, MA, USA).

Ogura S., Baldeosingh R., Bhutto I.A., Kambhampati S.P., McLeod D.S., Edwards M.M., Rais R., Schubert W, & Lutty G.A. (2020). A role for mast cells in geographic atrophy. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(8), 10117-10131.

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Antibody Characterization in Cell Research

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Calcium and magnesium free PBS (PBS; #10010‐023), and Hoechst 33258 (#H3569) were from Invitrogen (Waltham, MA). Triton X‐100 (#T8787) was obtained from Sigma–Aldrich (St. Louis, MO). Antibodies used in the current study were as follows: Mouse anti‐RPE65 (#NB100‐355) [clone 401.8B11.3D9]; Novus Biologicals, Centennial, CO), mouse anti‐DSG1 (#610273, BD Transduction, Franklin Lakes, NJ), mouse anti‐Cytokeratin 10 (#MA5‐13705 [clone DE‐K10]; ThermoFisher, Waltham, MA), rabbit anti‐Syntenin‐1 (#ab19903]; Abcam, Cambridge, MA), mouse anti‐Annexin II (#610068, BD Transduction Laboratories, Franklin Lakes, NJ), rat anti‐Integrin Beta 1 (AIIB2 clone, Developmental Studies Hybridoma Bank, Iowa City, IA), mouse anti‐Occludin (#66378‐1‐Ig, Proteintech, Rosemont, IL), rabbit anti‐Calreticulin (#12238 [clone D3E6]; Cell Signaling Technologies, Danvers, MA), anti‐SOD1 (#10269‐1‐AP, Proteintech), anti‐Catalase (#66765‐1‐Ig, Proteintech), anti‐MDA (#NBP2‐59367, Novus Biologicals), anti‐HSPA5 (#PA5‐19503, ThermoFisher), anti‐CLDN19 (#SC‐36597, Santa Cruz Technology, Dallas, TX), HRP‐conjugated donkey‐anti‐rat IgG (#712‐035‐153, Jackson ImmunoResearch Laboratories, West Grove, PA), HRP‐conjugated donkey‐anti‐mouse IgG (#715‐035‐150, Jackson ImmunoResearch) and HRP‐conjugated donkey‐anti‐rabbit IgG (#711‐035‐152, Jackson ImmunoResearch).

Hernandez B.J., Skiba N.P., Plössl K., Strain M., Liu Y., Grigsby D., Kelly U., Cady M.A., Manocha V., Maminishkis A., Watkins T., Miller S.S., Ashley‐Koch A., Stamer W.D., Weber B.H., Bowes Rickman C, & Klingeborn M. (2023). Polarized desmosome and hemidesmosome shedding via small extracellular vesicles is an early indicator of outer blood‐retina barrier dysfunction. Journal of Extracellular Biology, 2(10), e116.

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