Ab174309

Manufactured by Abcam

Sourced in United States

Ab174309 is a laboratory product offered by Abcam. It is a tool for use in scientific research applications. The core function of this product is to serve as a reagent or equipment for laboratory experiments and analyses, but no further details about its intended use or capabilities are provided.

Automatically generated - may contain errors

Lab products found in correlation

F4 80, by Thermo Fisher Scientific (1 mentions) Hoechst nuclear stain, by Merck Group (1 mentions) A1 confocal microscope, by Nikon (1 mentions) Nis element, by Nikon (1 mentions) Ab138501, by Abcam (1 mentions) Rabbit anti nf κb p65, by Cell Signaling Technology (1 mentions) Ba3677, by Boster Bio (1 mentions) Rabbit anti phospho nf κb p65, by Cell Signaling Technology (1 mentions) Sc 514414, by Santa Cruz Biotechnology (1 mentions) Mca1957, by Bio-Rad (1 mentions) Ab92377, by Abcam (1 mentions) Tcs nt confocal laser microscope, by Leica (1 mentions) Ab53444, by Abcam (1 mentions) Ab270449, by Abcam (1 mentions) Ab234437, by Abcam (1 mentions) Alexa fluor 488 or 594 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Mayer s hematoxylin, by Merck Group (1 mentions) A16792, by ABclonal (1 mentions) Sirius red, by Merck Group (1 mentions) Image pro plus 6, by Media Cybernetics (1 mentions)

7 protocols using ab174309

Immunohistochemical Analysis of Rev-Erbα

Check if the same lab product or an alternative is used in the 5 most similar protocols

Rev-Erbα was measured by incubation with anti-Rev-Erbα (Abcam, ab174309, rabbit anti-mouse, 1:200) and F4/80 (Thermo Fisher Scientific, Waltham, MA, Lot: 4339486 rat anti-mouse, 1:100) respectively, via manufacturer’s protocol for 1 h at room temperature, followed by a 1 h incubation at room temperature with 2 µg/mL goat anti-rabbit IgG conjugated with Cy5 (Jackson Immunoresearch) to be paired with the goat anti-rat IgG conjugated with Cy3 (Jackson Immunoresearch). A Hoechst nuclear stain (Sigma, B-2883) was applied at room temperature for 30 s followed by a single rinse of PBS to remove excess dye. Imaging conditions were maintained at identical settings with original gating performed using the negative control (no primary antibody). Images were taken from three random fields at 20 × 2 zoom with a Nikon A1 confocal microscope (purchased with 1S10OD019973-01 awarded to Dr. Simon C. Watkins). Quantification was performed using General Analysis Software on NIS Elements (Nikon).

Griepentrog J.E., Zhang X., Lewis A.J., Gianfrate G., Labiner H.E., Zou B., Xiong Z., Lee J.S, & Rosengart M.R. (2019). Frontline Science: Rev-Erbα links blue light with enhanced bacterial clearance and improved survival in murine Klebsiella pneumoniae pneumonia. Journal of leukocyte biology, 107(1), 11-25.

+ Open protocol

+ Expand

Protein Extraction and Western Blot Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

RIPA lysis buffer containing phosphorylase inhibitors, cocktails, and PMSF was used to digest tissue for protein extraction. Western blotting was performed as described previously [26 (link)]. The following primary antibodies were used: rabbit anti-NR1D1 (ab174309, Abcam), rabbit anti-NR1D1 (14506-1-AP, Proteintech), rabbit anti-tyrosine hydroxylase (TH) (25859-1-AP, Proteintech), rabbit anti-IBA1 (10904-1-AP, Proteintech), mouse anti-GFAP (60190-1-Ig, Proteintech), rabbit anti-NF-κB p65 (#8242, Cell Signaling Technology), rabbit anti-phospho-NF-κB p65 (#3033, Cell Signaling Technology), rabbit anti-NLRP3 (BA3677, BOSTER), mouse anti-ASC (sc-514414, Santa Cruz Biotechnology), rabbit anti-caspase-1 (22915-1-AP, Proteintech), rabbit anti-IL-1β (A16288, ABclone), rabbit anti-IL6 (A0286, ABclone), mouse anti-actin (66009-1-Ig, Proteintech), mouse anti-TNF Alpha (60291-1-Ig, Proteintech), rabbit anti-iNOS (22226-1-AP, Proteintech), rabbit anti-IL-18 (10663-1-AP, Proteintech), mouse anti-Arginase-1 (66129-1-Ig, Proteintech), rabbit anti-CD163 (16646-1-AP, Proteintech), rat anti-CD68 (MCA1957, BIO-RAD), Anti-Alpha-synuclein (ab138501, Abcam).

Kou L., Chi X., Sun Y., Han C., Wan F., Hu J., Yin S., Wu J., Li Y., Zhou Q., Zou W., Xiong N., Huang J., Xia Y, & Wang T. (2022). The circadian clock protein Rev-erbα provides neuroprotection and attenuates neuroinflammation against Parkinson’s disease via the microglial NLRP3 inflammasome. Journal of Neuroinflammation, 19, 133.

+ Open protocol

+ Expand

Immunofluorescence Analysis of Rev-erbα and Tff1

Check if the same lab product or an alternative is used in the 5 most similar protocols

Formalin-fixed paraffin-embedded tissues were cut int 4 μm sections which were deparaffinized with xylene and rehydrated through a graded series of alcohols. The tissue sections were placed in a repair box filled with citric acid antigen retrieval solution, and antigen retrieval was performed in a microwave oven. The slides were washed 3 times with PBS after natural cooling, 5 min each time and then, blocking was done with 3% BSA in PBS buffer for 30 min. Tissues were incubated with primary antibodies against Rev-erbα (ab174309, Abcam) and Tff1 (ab92377, Abcam) at 1 : 200 dilutions overnight at 4°C in humidified chambers followed by incubation with the corresponding secondary antibody for 60 min at room temperature in the dark. The slides were mounted with resin after dropping an appropriate amount of an antifluorescence quencher on the tissue. Fluorescence was detected with fluorescence microscopy. Fluorescence was detected by using a LEICA TCS NT laser confocal microscope.

Ke C., Xiaowen C., Yufeng W., Dongchang L., Huaqing Z, & Zhengguang W. (2022). Posttranslational Modifications of Rev-Erbα Protein and Abnormal Inflammatory Response in Gastric Cancer. Journal of Oncology, 2022, 6291656.

+ Open protocol

+ Expand

Evaluating NR1D1 and BMAL1 Expression in Cartilage

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunohistochemistry was performed to assess protein expression patterns in human and mouse cartilage using anti-NR1D1 antibody (Abcam ab174309, Cambridge, MA) and anti-BMAL1 antibody (Thermo Scientific PA1-523, Waltham, MA). Rabbit IgG (1 μg/ml) was used as a negative control in all experiments. For human cartilage, expression patterns were compared between normal and OA samples. In C57 BL/6 mice, we analyzed young normal and aged knees as a model of aging-related OA. We also analyzed knees from mice with surgically induced OA by destabilization of medial meniscus and medial collateral ligament resection ^{36 (link)}. The methods for tissue processing and immunohistochemistry were described earlier ^{36 (link)}.

Akagi R., Akatsu Y., Fisch K.M., Alvarez-Garcia O., Teramura T., Muramatsu Y., Saito M., Sasho T., Su A.I, & Lotz M.K. (2016). Dysregulated circadian rhythm pathway in human osteoarthritis: NR1D1 and BMAL1 suppression alters TGF-β signaling in chondrocytes. Osteoarthritis and cartilage, 25(6), 943-951.

+ Open protocol

+ Expand

Histological and Immunofluorescence Analysis of Carotid Arteries

Check if the same lab product or an alternative is used in the 5 most similar protocols

The carotid arteries specimens were embedded in the OCT compound and serially sectioned (5 μm). Hematoxylin and eosin (H&E), Masson's trichrome, and Sirius red staining were performed according to the manufacturers' protocols (Sigma). To evaluate the lipid content in the vasculature, sections were stained with oil red O, using Mayer's hematoxylin (Sigma) as the counter stain.
For immunofluorescence analysis, serial paraffin sections were incubated with different primary antibodies overnight at 4°C, followed by the corresponding fluorochrome-conjugated antibodies for detection. The following primary antibodies were used: rat monoclonal antibody against CD68 (Abcam, ab53444), rabbit monoclonal antibody against NLRP3 (Abcam, ab270449), rabbit monoclonal antibody against IL-1β (Abcam, ab234437), rabbit polyclonal antibody against caspase-1 (ABclonal, A16792), and rabbit monoclonal antibody against NR1D1 (Abcam, ab174309). After they reacted with the corresponding Alexa Fluor 488 or 594-conjugated secondary antibodies (Invitrogen), DAPI was added to visualize nuclei. Fluorescent images were acquired with a laser scanning confocal microscope (Leica TCS SP8, Leica Microsystems). All the images and staining intensities were acquired and measured with Image-Pro Plus 6.0 (Media Cybernetics Inc.).

Wu Z., Liao F., Luo G., Qian Y., He X., Xu W., Ding S, & Pu J. (2021). NR1D1 Deletion Induces Rupture-Prone Vulnerable Plaques by Regulating Macrophage Pyroptosis via the NF-κB/NLRP3 Inflammasome Pathway. Oxidative Medicine and Cellular Longevity, 2021, 5217572.

+ Open protocol

+ Expand

Klebsiella pneumoniae Growth Kinetics and Interventions

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Klebsiella pneumoniae strain 43816, serotype 2 (American Type Culture Collection, Manassas, VA) was grown in tryptic soy broth overnight at 37°C. A total of 200 µL of this overnight culture was inoculated into fresh tryptic soy broth and grown for 1.5 h. A standard growth curve of bacterial culture measured by absorbance at 600 nm was generated to determine mid-log phase of growth as we previously have done.^{15 (link),16 (link)} Inoculum concentration, measured in CFU, was determined by serial 10-fold dilutions of bacteria plated on tryptic soy agar plates. Bacteria were resuspended in PBS before use.
Antibodies for Rev-Erbα (ab174309) and tubulin (ab4074) were obtained from Abcam (Cambridge, MA). α-Bungarotoxin was obtained from Tocris (21331), dissolved in deionized water and administered i.p. at a dose concentration of 1 µg/kg. The Rev-Erbα agonist SR9009 (EMD Millipore, Billerica, MA) was dissolved in vehicle (15% Kollifor) and administered i.p. at a dose concentration of 100 mg/kg.

+ Open protocol

+ Expand

Immunofluorescence Analysis of Cellular Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

GEFs of the third generation were subjected to immunofluorescence staining as previously described (24 (link), 33 (link)). Briefly, the GEFs were fixed in 4% paraformaldehyde at 25°C for 20 min, permeabilized with 0.1% Triton X-100 for 30 min, and subsequently blocked for 30 min with 5% bovine serum albumin at 25°C. The GEFs were then incubated with anti-vimentin (a marker of stromal cells, Abcam, Cambridge, MA, USA, ab8978, 1:100), anti-cytokeratin (a marker of epithelial cells, Abcam, ab181597, 1:100), anti-BMAL1 (Abcam, ab93806, 1:100), or anti-NR1D1 (Abcam, ab174309, 1:100) antibodies diluted in PBS at 4°C for 24 h. After washing, the cells were treated with PBS containing fluorescein-conjugated secondary antibody (1:200, Thermo Fisher, Shanghai, China) and DAPI (blue), and the immunostaining signal was detected using a fluorescence microscope (Nikon, Tokyo, Japan).

Gao D., Zhao H., Dong H., Li Y., Zhang J., Zhang H., Zhang Y., Jiang H., Wang X., Wang A., Jin Y, & Chen H. (2022). Transcriptional Feedback Loops in the Caprine Circadian Clock System. Frontiers in Veterinary Science, 9, 814562.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!