Cholesterol (K1805006), Span 20 (B1806043), and Span 60 (S112961) were obtained from Aladdin company (Shanghai, China). Carbopol powder (0101536985) was purchased from Chineway (Shanghai, China). Cel (19041101) was supplied by Chengdu Pufei De Biotech Co., Ltd. (Chengdu, China). Hair remover spray foam was purchased from Linco Care Ltd. (Carrington, M, USA). IMQ cream (5%) was obtained from Aldara (Health Care Limited, Loughborough, UK). IMQ powder was acquired from MedChemExpress (Shanghai, China). Protopic (0.1%, tacrolimus) was supplied by Kangaiduo Drug Store (Guangzhou, China). Absolute ethanol, chloroform, acetonitrile, methanol, and formic acid were from Merck (Darmstadt, Germany). Milli-Q water was produced using a Millipore Direct-Q ultrapure water system (Millipore, Bedford, USA). Goat serum was from Boster Systems, Inc. (AR0009; Pleasanton, CA, USA). Fetal bovine serum (FBS), IFN-β (PA5-20390), and ProLong™ Gold Antifade Mountant with DAPI (P36935) were acquired from Thermo Fisher Scientific (Waltham, MA, USA). Ki-67 (ab15580), IL-6 (ab179570), TNF-α (ab8348), goat anti-mouse IgG H&L (ab150115) and goat anti-rabbit IgG H&L (ab150077) were purchased from Abcam (Cambridge, UK). Paraformaldehyde (4%) was obtained from Phygene Life Sciences Co., Ltd. (Fuzhou, China). 1, 1ʹ-Dioctadecyl-3,3,3ʹ,3ʹ-tetramethylindocarbocyanine (DiI) was from Invitrogen (Carlsbad, CA, USA).
Goat anti rabbit igg h l ab150077
Manufactured by Abcam
Sourced in United Kingdom, China
Goat anti-rabbit IgG H&L (ab150077) is a secondary antibody product that binds to the heavy and light chains of rabbit immunoglobulin G (IgG). It is designed for use in various immunoassays and other applications where detection of rabbit IgG is required.
Automatically generated - may contain errors
Lab products found in correlation
Direct q ultra pure water system, by Merck Group (1 mentions)
Imq powder, by MedChemExpress (1 mentions)
Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Chloroform, by Merck Group (1 mentions)
Formic acid, by Merck Group (1 mentions)
Absolute ethanol, by Merck Group (1 mentions)
Ab181602, by Abcam (1 mentions)
Novex ecl chemiluminescent substrate reagent kit, by Thermo Fisher Scientific (1 mentions)
Polyvinylidene difluoride pvdf membrane, by Merck Group (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Ripa lysate, by Beyotime (1 mentions)
Chemidoc touch imaging system, by Bio-Rad (1 mentions)
Beyoecl plus, by Beyotime (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Anti neun antibody ab177487, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride, by Merck Group (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
5 protocols using goat anti rabbit igg h l ab150077
1
Topical Formulation for Psoriasis Treatment
2
Western Blot Analysis of TRIM14 Protein
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BC cells were lysed with radioimmunoprecipitation assay (RIPA) lysate (Beyotime, Shanghai, China) and the supernatant was immediately obtained by high-speed centrifugation to extract total protein, with the total protein concentration examined by a BCA protein assay kit (Beyotime, Shanghai, China). The protein was subsequently heated at 100°C for 10 min. Specifically, proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA), which were subsequently blocked with 5% skimmed milk for 1 h and rinsed three times with Tris Buffered Saline with Tween 20 (TBST). Proteins were subsequently incubated with primary anti-TRIM14 antibody (PA5-50806, 1:1000, Thermo Fisher Scientific) and anti-GAPDH antibody (ab181602, 1:1500, Abcam, Shanghai, China) overnight at 4°C. After the membrane was rinsed by TBST again, the PVDF membrane was incubated with Goat Anti-Rabbit IgG H&L (ab150077, 1:5000, Abcam, Shanghai, China) for 1 h at room temperature. Ultimately, proteins were specifically visualized by a Novex™ ECL Chemiluminescent Substrate Reagent Kit (Thermo Fisher Scientific), with GAPDH as the internal reference.
3
Western Blot Analysis of Autophagy Markers in Jejunum
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Relative protein levels of Beclin1, Sequestosome 1 (P62 /SQSTM1, P62), LC3, and β-actin in the jejunum were determined by Western blotting, as described previously [5 (link)]. The primary antibodies used in the present study were as follows: anti-Beclin1 (#3495), anti-P62 (#23214), anti-LC3B, and anti-β-actin (#4970) (Cell Signaling Technology Co., Ltd., Danvers, MA, USA). The second antibody, Goat Anti-Rabbit IgG H&L (ab150077), was purchased from Abcam (Shanghai, China). Chemiluminescent reagent (BeyoECL Plus, Beyotime, Shanghai, China) with a ChemiDoc™ Touch Imaging System (Bio-Rad, Philadelphia, PA, USA) was used to visualize the bands of the protein. The resultant signals were quantified as described previously [25 (link)].
4
Immunofluorescence Assay of Kir4.1 Expression
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Immunofluorescence was determined in vitro and ex vivo. In vitro, cells were cultured at 33°C or 39°C for 3 or 7 days. Cells were then seeded (4 × 104 cells/ml) in 4-well dishes. After culturing for 24 hours, each sample was washed with 1 ml PBS, and 500 μl of 4% paraformaldehyde was added to each well and fixation proceeded for 1 hour. The paraformaldehyde was removed, and each sample was washed three times for 5 minutes each in PBS. After the final wash, 500 μl of 0.1% Triton X-100 was added and incubated for 15 minutes at room temperature. Triton X-100 was removed, and samples were washed three times for 5 minutes each in PBS. The primary antibody (Kir4.1, APC-035; Alomone labs, Jerusalem, Israel) diluted 1 : 100 in 5% normal goat serum (NGS) was added, and the cells were incubated for 1 hour at room temperature. PBS (500 μl) was then added to each well and washed three times (5 minutes per wash). The secondary antibody (goat anti-rabbit IgG H&L, ab150077; Abcam, Cambridge, UK) was diluted 1 : 200 in 5% normal goat serum (NGS) and incubated for 1 hour at room temperature. Each sample was then mounted using mounting solution and stained with 4′,6-diamidino-2-phenylindole (DAPI), and a cover glass was added. The solution was dried for 24 hours in a dark room and examined using confocal microscopy.
5
Neuronal Differentiation Assessment of SK-N-SH Cells
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Neuronal differentiation of SK-N-SH cells was determined using the neuronal marker, NeuN. Cultured cells were washed three times with D-PBS-free and then fixed with 4% paraformaldehyde in D-PBS-free (pH 7.4) for 10 min. The samples were washed twice with D-PBS-free and then treated with 0.2% TritonX-100 in D-PBS-free for 10 min at 25 °C. Treated samples were washed twice in D-PBS-free for 5 min and incubated with 0.5% BSA in D-PBS-free for 30 min at 25 °C. After removing the 0.5% BSA in D-PBS-free, the treated samples were incubated with 1000-fold diluted primary antibody (Anti-NeuN antibody, ab177487; Abcam plc, Cambridge, UK) in 0.5% BSA in D-PBS-free, overnight at 4 °C. The treated samples were washed twice with 0.5% BSA in PBS for 5 min and then incubated with 4000-fold diluted secondary antibody (Goat Anti-Rabbit IgG H&L, ab150077; Abcam plc) and 1 μg/mL DAPI (4′,6-Diamidino-2-phenylindole, dihydrochloride; Sigma–Aldrich Co. LLC, St. Louis, MO, USA) in 0.5% BSA in D-PBS-free for 1 h at room temperature in the dark. Treated samples were washed twice with 0.5% BSA in D-PBS-free for 5 min, and the expression of NeuN protein in SK-N-SH cells was observed using a fluorescence microscope (Olympus Co., Tokyo, Japan).
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