Cells were fixed with 4% formaldehyde (BioSharp, Hefei, China) for 10 min at room temperature. Cells were then permeabilized with 0.5% Triton X-100 (BioSharp, Hefei, China) for 2 min and treated with a blocking serum (Solarbio, Beijing, China) for more than 5 h. Primary antibody, anti-COUP TF1 (sc-74560, 1:50, Santa Cruz, TX, USA), and anti-Ki67 (ab15580, 1:200, Abcam, Cambridge, UK) were incubated overnight for immunofluorescence. After being washed to remove unbound primary antibodies, cells were incubated with the secondary antibody Dnk pAb to Rb IgG (Alexa Fluor® 488) (ab150074, 1:1000, Abcam, UK), donkey anti-mouse IgG H&L (Alexa Fluor 594) (ab15010B, 1:1000, Abcam, UK), and DAPI (Biosharp, Hefei, China) for 2 h under dark conditions. Images were acquired with a Leica SP8 (Wetzlar, Germany) confocal microscope.
Ab150074
Manufactured by Abcam
Sourced in United Kingdom, United States, China
Ab150074 is a recombinant protein that binds to a specific target. Its core function is to serve as a research tool for target detection and quantification. Detailed technical specifications are available upon request.
Automatically generated - may contain errors
Lab products found in correlation
Ab150105, by Abcam (10 mentions)
Ab150113, by Abcam (4 mentions)
Ab150129, by Abcam (3 mentions)
Ab150073, by Abcam (3 mentions)
Ab15580, by Abcam (2 mentions)
Ab3526, by Abcam (2 mentions)
Ab13970, by Abcam (2 mentions)
Ab34710, by Abcam (2 mentions)
Efficient navigation zen , by Zeiss (2 mentions)
H 1200, by Vector Laboratories (2 mentions)
4 6 diamindino 2 phenyindole dapi, by Merck Group (2 mentions)
Ab33168, by Abcam (2 mentions)
Paraformaldehyde (pfa), by Thermo Fisher Scientific (2 mentions)
Lsm800 confocal microscope, by Zeiss (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Lsm 700, by Zeiss (2 mentions)
Formaldehyde, by Biosharp (1 mentions)
Triton x 100, by Biosharp (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Biosharp (1 mentions)
Blocking serum, by Solarbio (1 mentions)
41 protocols using ab150074
1
Immunofluorescence Staining of COUP TF1 and Ki67
2
Immunofluorescence Staining of Paraffin Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All tissue samples were fixed and processed into paraffin blocks. After dewaxing, antigen repair (10 mmol/L sodium citrate for 5 min in a pressure cooker) and removing endogenous enzymes in Hydrogen Peroxide Block, the sections were incubated with the primary antibodies anti-GFP (ab13970, Abcam, UK, the dilution ratio was 1:500) and anti-Perilipin (ab3526, Abcam, UK, the dilution ratio was 1:200) at 4ºC overnight. Then the samples were incubated in secondary antibody (703-545-155, Jackson ImmunoResearch, USA; ab150074, Abcam, UK, the dilution ratio was 1:500) for 1-2 h at room temperature in the dark. After rinsing three times in 1X PBS for 5 min each, the slides were counterstained with Hoechst (C1026, Beyotime, China, the dilution ratio was 1:500) to reveal cell nuclei. Finally, fluorescence microscopy was used to evaluate specimen.
3
Quantitative Proinsulin and Insulin Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Corresponding reagents were prepared and placed at room temperature. Sample or standard solution (100 µl) was added in the sample wells or standard wells, respectively. Corresponding antibodies, including proinsulin antibody (1:500; cat. no. ab8304; Abcam, Cambridge, MA, USA) and insulin antibody (1:500; cat. no. ab100578; Abcam) were then added for incubation at room temperature for a total of 2 h. Subsequently, horseradish peroxidase-labeled antibodies were added (1:1,000; cat. no. ab150074; Abcam). After incubation for 1 h at room temperature, 100 µl substrate was added, followed by colour reaction in the dark for 10 min. The absorbance at the wavelength of 450 nm was detected by a microplate reader (Bio-Rad Laboratories, Hercules, CA, USA).
4
Immunofluorescence Analysis of γH2AX in Tissue Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Indicated de-paraffinized tissue sections were used for immunofluorescence staining. Antigen retrieval was performed by incubating them in Retrieval buffer-A (10 mM Tris-Cl pH-9.0, 1 mM EDTA, and 0.05% Tween-20) for 20 min at room temperature (RT), followed by permeabilization with 0.1% saponin for 30 min at RT. These permeabilized sections were then blocked and incubated with the ɤH2AX-antibody (Cell signaling technology #9718) in blocking buffer (10% goat serum in TBS-0.2% Triton-X100) at 4 °C overnight. Fluorochrome conjugated anti-Rabbit (Alexa-555; Abcam Ab150074) antibody was used as the secondary antibody. The negative control staining was performed simultaneously with ɤH2AX staining. Stained sections were then imaged by using a CCD camera connected to an inverted fluorescence microscope (Cell Observer, Carl Zeiss, GmbH, Göttingen, Germany), using an ×63 oil objective. Images were further processed using ImageJ (Fiji).
5
Paraffin-embedded Immunofluorescence Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All tissue samples were fixed and processed into paraffin blocks. After dewaxing, antigen repair (10 mmol l−1 sodium citrate for 5 min in a pressure cooker) and removing endogenous enzymes in hydrogen peroxide block, the sections were incubated with the primary antibodies anti-GFP (ab13970, Abcam, UK, the dilution ratio was 1:500) and anti-Perilipin (ab3526, Abcam, UK, the dilution ratio was 1:200) at 4 °C overnight. Then the samples were incubated in secondary antibody (703–545–155, Jackson ImmunoResearch, USA; ab150074, Abcam, UK, the dilution ratio was 1:500) for 2 h at room temperature in the dark. After rinsing three times in 1× PBS for 5 min each, the slides were counterstained with Hoechst (C1026, Beyotime, China, the dilution ratio was 1:500) to reveal cell nuclei. Finally, fluorescence microscopy was used to evaluate specimen.
6
Immunofluorescent Staining of PTX3 in Paraffin-Embedded Tumor Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Paraffin sections were deparaffinized, rehydrated and antigen retrieval was performed in sodium citrate. The immunofluorescent staining protocol was identical to the one used for spheroids. Tumor slides were incubated with a polyclonal anti-PTX3 (#HPA069320 Atlas Antibodies Ozyme, St Quentin, France) and then with an Alexafluor 555-labelled donkey anti-rabbit (#ab150074 Abcam). Cell nuclei were counterstained with 1 μg/mL DAPI (Sigma Aldrich). Slides were visualized using a Zeiss Axio-Imager Z2 fluorescence microscope at 20× magnification.
7
Experimental Autoimmune Encephalomyelitis Induction and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rat MOG 35–55 peptide (MEVGWYRSPFSRVVHLYRNGK; >95% purity) was synthesized by Xi’an Biotechnology Co., (Xi’an, China). Heat-killed Mycobacterium tuberculosis H37Ra was obtained from Difco (USA). Complete Freund's adjuvant (CFA) was obtained from Sigma Aldrich (USA), and pertussis toxin (PTX) from Alexis (Germany). The following antibodies were used in this study: FITC-anti-mouse CD45 (eBioscience, USA), PE-Cyanine5-anti-mouse CD117 (eBioscience), anti-mouse IL-9 (BE0181; BioXCell, USA), anti-mouse IgG2a isotype control (BE0085; BioXCell), anti-mouse IL-9 receptor (IL-9R) (SC699; Santa Cruz, USA), anti-mouse IL-2Rγ (SC668; Santa Cruz), anti-mouse IgG isotype control (GTX35009; Santa Cruz), and donkey pAb to Rb IgG Alexa Flour 555 (Ab150074; Abcam, USA).
8
Multimodal Imaging of Liver Fibrosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The liver tissues were embedded in OCT and stored at -80°C. OCT-embedded liver tissues were cut into 6μm sections. The sections were fixed in 4% paraformaldehyde for 15 min and then washed with PBS. Next, the sections were incubated in permeabilization solution (0.3% Triton X-100 in PBS) for 15 min and then washed with PBS. Then, the sections were incubated with anti-collagen I (ab34710, Abcam), anti-Ki67 (ab15580, Abcam) or α-SMA (ab7817, Abcam) antibodies. After washing, the sections were incubated with Alexa Fluor 488-conjugated donkey anti-rabbit IgG (711-545-152, Jackson ImmunoResearch Labs), Alexa Fluor 555-conjugated donkey anti-rabbit IgG (ab150074, Abcam) or Alexa Fluor 594-conjugated donkey anti-mouse IgG (715-585-150, Jackson ImmunoResearch Labs). The sections were washed with PBS, counterstained with 4,6-diamidino-2-phenylindole (DAPI) (10236276001, Roche) and mounted with a cover glass. The images were captured with a confocal laser microscope setup (LSM980, Zeiss) and processed using ZEN (Zeiss).
9
Immunofluorescence Analysis of β-catenin, LGR6, and hnRNPK
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed, permeabilized with 0.5% Triton X-100 (Beyotime, Shanghai, China), and blocked with 5% BSA. Primary antibodies specific to β-catenin (dilution 1:200, EM0306, Huabio, Hangzhou, China), LGR6 (dilution 1:200, ab126747, Abcam, Shanghai, China) and HnRNPK (dilution 1:200, SC60-03, Huabio, Hangzhou, China) were then added and incubated overnight at 4 °C. Secondary antibodies labeled with Cy3 (A0521, Beyotime, Shanghai, China) or Alexa Fluor 555 (ab150074, Abcam, Shanghai, China) were used to incubate the samples for 2 h. DAPI (S2110, Solarbio, Beijing, China) and phalloidine (6 μmol/L, Invitrogen, Carlsbad, CA, USA) were applied to stain the nuclei and cytoskeleton. Finally, cell samples were observed with CLSM.
10
Immunofluorescence Analysis of Periostin and Collagen III
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!