Goat anti mouse albumin antibody

Manufactured by Fortis Life Sciences

Sourced in United States

Goat anti-mouse albumin antibody is a laboratory reagent used for the detection and quantification of mouse albumin in various biological samples. It is a polyclonal antibody raised in goats against mouse albumin protein.

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Lab products found in correlation

Sk 4100, by Vector Laboratories (1 mentions) Bx41 microscope, by Olympus (1 mentions) Streptavidin alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Isolectin gs ib4, by Merck Group (1 mentions) Te ez c1 confocal system, by Nikon (1 mentions) Nis elements software, by Nikon (1 mentions) 3 3 5 5 tetramethylbenzidine (tmb), by Merck Group (1 mentions) Benchmark plus microplate spectrophotometer, by Bio-Rad (1 mentions) Creatinine colorimetric assay kit, by Cayman Chemical (1 mentions) Polyoxyethylene octyl phenyl ether, by Fujifilm (1 mentions) Ipgel, by Nippon Genetics (1 mentions) Target retrieval solution, by Agilent Technologies (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Vectashield mounting medium with dapi, by Vector Laboratories (1 mentions) Serotonin elisa kit, by Enzo Life Sciences (1 mentions) Mouse tnf alpha duoset elisa, by R&D Systems (1 mentions) Evos fl auto 2 cell imaging system, by Thermo Fisher Scientific (1 mentions) Glucose assay kit, by Abcam (1 mentions) Ultra sensitive mouse insulin elisa kit, by Crystal Chem (1 mentions)

7 protocols using goat anti mouse albumin antibody

Immunohistochemical Analysis of Albumin Expression After SAH

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Immunohistochemistry was performed as described in a previous study^{37 (link)}. Rats were euthanized 72 h after SAH induction and transcardially perfused with 0.1
mol/l PBS (pH 7.4) followed by 4% paraformaldehyde (pH 7.4) as described above. Then, the
brains were removed, fixed, dehydrated, and cut into coronal frozen sections (7 μm). The
sections were incubated with a goat anti-mouse albumin antibody (1:500, Bethyl
Laboratories, Cat# A90-134A) at 25°C for 2 h and washed with PBS (pH 7.4). After the
sections were further incubated with secondary antibody for an additional 1 h at 25°C, the
reaction product was visualized using a DAB peroxidase substrate kit SK-4100 (Vector
Laboratories, Burlingame, CA, USA). An Olympus BX41 microscope was used to capture the
images.

Chen J., Jin H., Xu H., Peng Y., Jie L., Xu D., Chen L., Li T., Fan L., He P., Ying G., Gu C., Wang C., Wang L, & Chen G. (2019). The Neuroprotective Effects of Necrostatin-1 on Subarachnoid Hemorrhage in Rats Are Possibly Mediated by Preventing Blood–Brain Barrier Disruption and RIP3-Mediated Necroptosis. Cell Transplantation, 28(11), 1358-1372.

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Assessing Blood-Retinal Barrier Integrity

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Assessment of the BRB was conducted in cryosections, by assessing albumin leakage from the retinal vasculature. The vasculature was visualised using biotinylated isolectin GS-IB4 (Sigma) followed by binding to streptavidin–Alexa Fluor 488 (Life Technologies, Paisley, UK). Albumin was localised using a goat anti-mouse albumin antibody (Bethyl Laboratories, Montgomery, TX, USA) followed with anti-goat Alexa Fluor 568 (1:500; Life Technologies). Vessel or neuropile-localised fluorescence was visualised by using a Nikon TE EZ-C1 confocal system (Nikon, Kingston Upon Thames, UK). Using NIS Elements software (Nikon) a threshold value of 12,000 was set for measuring image intensity/brightness. The area (μm²) of the albumin staining was measured with the 12,000 brightness intensity threshold in the retinal sections. Images were taken at three separate points on the central retina at magnification ×40 and n = 5 per group of mice.

McVicar C.M., Ward M., Colhoun L.M., Guduric-Fuchs J., Bierhaus A., Fleming T., Schlotterer A., Kolibabka M., Hammes H.P., Chen M, & Stitt A.W. (2015). Role of the receptor for advanced glycation endproducts (RAGE) in retinal vasodegenerative pathology during diabetes in mice. Diabetologia, 58(5), 1129-1137.

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Quantifying Urinary Albumin Levels

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Albumin concentration in the urine was determined using the following ELISA procedure. 96-wells plates were coated overnight with a goat anti-mouse albumin antibody (BETHYL Laboratories, Inc. Montgomery, Texas, USA) diluted 1:100 in 0.05 M Carbonate–bicarbonate at pH 9.6. After blocking with 50 mM Tris, 0.14 M NaCl, 1% BSA, pH 8.0, urinary samples were incubated for 1 h in various dilutions. Bound albumin was detected using an HRP conjugated goat anti-mouse albumin antibody diluted 1:75,000 (BETHYL Laboratories, Inc. Montgomery, Texas, USA). Binding was visualized using 3,3′,5,5′- tetramethylbenzidine (Sigma-Aldrich, Zwijndrecht, The Netherlands) where after 1 M H₂SO₄ was added and absorbance was measured at 450 nm (Benchmark Plus microplate spectrophotometer, BIORAD). Urinary albumin concentration was determined using a standard curve with known albumin concentrations.
Urinary creatinine was measured using the creatinine colorimetric assay kit (Cayman Chemicals, Ann Arbor, Michigan, USA) according to manufacturer’s instructions. Albumin values were adjusted for creatinine values by dividing the urinary albumin concentration by the urinary creatinine concentration resulting in the albumin/creatinine ratio (ACR).

Talsma D.T., Katta K., Ettema M.A., Kel B., Kusche-Gullberg M., Daha M.R., Stegeman C.A., van den Born J, & Wang L. (2018). Endothelial heparan sulfate deficiency reduces inflammation and fibrosis in murine diabetic nephropathy. Laboratory investigation; a journal of technical methods and pathology, 98(4), 427-438.

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Immunocytochemistry of Liver Cells

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The cultured cells were fixed and stained as previously described49 (link). Primary antibodies were as follows; mouse anti-human Hnf4a (PPMX), goat anti-human Foxa3 (Santa Cruz), goat anti-mouse albumin antibody (Bethyl Laboratories), mouse anti-mouse E-cadherin (Abcam), mouse anti-mouse cytchromeP450 1A2 (Abcam) at 1:200 respectively. The sections were incubated for two hours with secondary antibodies; Alexa 488-conjugated rabbit anti-goat IgG or goat anti-mouse IgG at 1:1000. After washing, the stained sections were covered with Vectashield mounting medium with DAPI (Vector Laboratories).
Spheroids were first embedded in iPGel (NIPPON Genetics) and fixed with 4% PFA overnight, then embedded in paraffin and cut in 5 μm thick. After deparaffinization and antigen retrieval using a Target Retrieval Solution (Dako), nonspecific binding was blocked with 1% bovine serum albumin (Sigma Aldrich) dissolved in 0.1% Polyoxyethylene Octylphenyl Ether (Wako) in PBS. The primary and secondary antibodies reactions were carried out in the same way for cultured cells.

Katayama H., Yasuchika K., Miyauchi Y., Kojima H., Yamaoka R., Kawai T., Yukie Yoshitoshi E., Ogiso S., Kita S., Yasuda K., Sasaki N., Fukumitsu K., Komori J., Ishii T, & Uemoto S. (2017). Generation of non-viral, transgene-free hepatocyte like cells with piggyBac transposon. Scientific Reports, 7, 44498.

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Platelet Inhibition and Hemostasis

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The effect of platelet inhibition by ticagrelor was assessed by bleeding volume and plasma concentrations of PF4 and serotonin. For the bleeding volume, 2 mm of the tail tip was amputated and blood was captured in a 15 mL tube with saline. The tube was centrifuged twice to pellet the erythrocytes (1550 g, 5 minutes), the erythrocytes were lysed with 5 mL of demineralized water and the optical density of the lysates was determined by spectrophotometer at 570 nm. Plasma PF4 and serotonin were measured by ELISA in citrated plasma at the end of the study, with mouse CXCL4/PF4 ELISA kit (R&D Systems, Minneapolis, MN, USA) and Serotonin ELISA kit (Enzo Life Sciences, New York, NY, USA), respectively.
Plasma concentrations of creatinine, urea, ALAT, and ASAT were determined using standardized clinical diagnostic protocols. Albumin levels in 24-hours urine were measured with an enzyme-linked immunosorbent assay using the goat anti-mouse albumin antibody (Bethyl Laboratories, Montgomery, TX, USA). TNF-α in plasma and kidney homogenates was measured with the mouse TNF-alpha DuoSet ELISA (R&D Systems, Minneapolis, MN, USA).

Uil M., Butter L.M., Claessen N., Larsen P.W., Florquin S, & Roelofs J.J.T.H. (2020). Platelet inhibition by ticagrelor is protective against diabetic nephropathy in mice. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(10).

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Histopathological Assessment of Choroid Plexus

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H&E-stained sections were scored, using a range from 0 to 4, based on stromal expansion and cellular infiltration in the choroid plexus of the third ventricle, as described.^{26 (link)}Coronal sections of the paraffin-embedded brain tissue were cut for immunofluorescent staining. All sections were blocked initially with 20% normal horse serum. Sections were incubated with a primary goat antimouse albumin antibody (1:300; Bethyl, Montgomery, TX), followed by a secondary Cy5-conjugated donkey antigoat antibody (1:500; Jackson ImmunoResearch, West Grove, PA), for albumin staining. IgG deposition in the brain was assessed by staining with a directly Cy5-conjugated donkey antimouse IgG (1:100; Jackson ImmunoResearch). Leukocyte infiltration was also assessed by staining for CD3 and B220, using primary antibodies of rabbit antimouse (1:100; Invitrogen, Carlsbad, CA) and rat antimouse (1:100; BD Biosciences, San Jose, CA) and secondary antibodies of Cy5-conjugated donkey antirabbit and AF488-conjugated donkey antirat (1:100; Jackson ImmunoResearch), respectively. The secondary antibody applied alone served as a negative control.
Fluorescence-stained slides were imaged under an Invitrogen microscope (Invitrogen EVOS FL Auto 2 Cell Imaging System). Images were analyzed and processed using ImageJ (National Institutes of Health, Bethesda, MD).

Huang M., Stock A., Mike E., Herlitz L., Kolbeck R, & Putterman C. (2019). Anti-IFNAR treatment does not reverse neuropsychiatric disease in MRL/lpr lupus mice. Lupus, 28(13), 1510-1523.

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Standardized Biomarker Evaluation Protocol

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Plasma creatinine and plasma urea were measured using standardized clinical diagnostic protocols. Albumin levels in 24-hour urine were measured by means of sandwich ELISA using the goat anti-mouse albumin antibody (Bethyl Laboratories, Montgomery, TX, USA). Urinary glucose concentrations were determined in deproteinized samples (PCA kit, Biovision, Milpitas, CA, USA) by a glucose assay kit (Abcam, Cambridge, UK) and fasting insulin levels were determined by ultrasensitive mouse insulin ELISA kit (CrystalChem, Downers Grove, USA). Insulin resistance was estimated with the homeostatic model assessment- insulin resistance (HOMA-IR) using the following formula: HOMA-IR = [fasting plasma insulin(µU/mL)*fasting blood glucose (mmol/L)/22.5]^{50 (link),51 (link)}. HOMA-beta was calculated as follows: HOMA-beta = (20*fasting plasma insulin)/(fasting blood glucose-3,5)^{51 (link)}.

Uil M., Scantlebery A.M., Butter L.M., Larsen P.W., de Boer O.J., Leemans J.C., Florquin S, & Roelofs J.J. (2018). Combining streptozotocin and unilateral nephrectomy is an effective method for inducing experimental diabetic nephropathy in the ‘resistant’ C57Bl/6J mouse strain. Scientific Reports, 8, 5542.

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