Nf κb p65 f 6

Manufactured by Santa Cruz Biotechnology

NF-κB p65 [F-6]: is a lab equipment product used for the detection and analysis of the NF-κB p65 subunit. It is a reliable tool for researchers studying the NF-κB signaling pathway and its role in various cellular processes.

Automatically generated - may contain errors

Lab products found in correlation

Ecl plus, by Thermo Fisher Scientific (3 mentions) Rabbit anti rat hrp, by Agilent Technologies (1 mentions) Mca2388, by Bio-Rad (1 mentions) Alexa fluor 594 goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Goat anti mouse igg hrp, by Santa Cruz Biotechnology (1 mentions) Vinculin antibody 7f9, by Santa Cruz Biotechnology (1 mentions) Goat anti rb igg hrp, by Santa Cruz Biotechnology (1 mentions) Ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions) Gfp b 2, by Santa Cruz Biotechnology (1 mentions) Twist 2c1a, by Santa Cruz Biotechnology (1 mentions) Bcl 2 c 2, by Santa Cruz Biotechnology (1 mentions) β actin c4, by Santa Cruz Biotechnology (1 mentions) Image lab 4.1 analysis software 4, by Bio-Rad (1 mentions) Gel imaging system, by Bio-Rad (1 mentions) Inverted confocal microscope, by Olympus (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)

6 protocols using nf κb p65 f 6

Wound Healing Efficacy of MASE Gel

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

Twenty male
Rattus norvegicus(Wistar) rats were randomly divided into four groups. Their left buccal mucosae were injured by means of a punch biopsy with a 6 mm diameter and 1 mm depth after being placed under general anesthesia ether.
5
The wound was ensured to be confined to the epithelium, without damaging the underlying muscle. Each of the four groups was then treated with MASE gel of varying concentrations three times a day (every 6–8 h); 0% (as control), MASE 25%, MASE 37.5%, and MASE 50%.
After 3 days of application, the
Rattus norvegicuswere sacrificed with lethal dose of ether by inhalation and their buccal mucosae biopsied for immunohistochemistry (IHC) analysis. Immunohistochemical staining was conducted using anti-mouse TNF-α monoclonal antibodies (Santa Cruz Biotechnology Inc., TNF-α [M-18]: sc 1348) and NF-κB monoclonal antibodies (Santa Cruz Biotechnology Inc.; NF-κB p65 [F-6]: sc 8008). The number of macrophage cells showing the expressions of TNF-α and NF-κB was calculated in three different field locations using a light microscope (Olympus, United States) at 400× magnification, and subsequently averaged.

Apriasari M.L., Pramitha S.R., Puspitasari D, & Ernawati D.S. (2020). Anti-Inflammatory Effect of Musa acuminata Stem. European Journal of Dentistry, 14(2), 294-298.

+ Open protocol

+ Expand

Analyzing Muscle Fiber Integrity Using Acid Phosphatase Staining

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

Acid phosphatase staining was analysed for compromised muscle fibre integrity by incubating TA cryosections in acid phosphatase buffer (HPR reagent, 0.1 M acetate buffer pH 5.0, 50 mg/mL naphthol AS-BI phosphate) for 90 min at 37 °C before being washed and counterstained with a 1:30 dilution of Harris haematoxylin for 1 min. The stained slides were mounted using hydromount.
Immunocytochemistry was performed as previously described [42 (link)] with the following antibodies: monoclonal anti-mouse Pax7 (DSHB-Pax7) (1:1), monoclonal anti-mouse Myosin heavy chain 3 (DSHB-F1.652) (1:1), Rat anti-mouse CD31 (BioRad MCA2388) (1:150), Rat anti-mouse F4-80 (BioRad MCA497R) (1:200), polyclonal rabbit anti-MyoD (Santa Cruz sc-760) (1:200), monoclonal mouse anti-CD68 (Abcam ab955) (1:200), and monoclonal mouse anti-NF-κB-P65 (F-6) (Santa Cruz sc8008) (1:200). Secondary antibodies used were Alexa Fluor 488 Goat anti-mouse IgG (Invitrogen A11029) (1:200), Alexa Fluor 594 Goat anti-Rabbit IgG (Invitrogen A11012) (1:200), and Rabbit anti-Rat HRP (DAKO P0450) (1:200). Sections were mounted using mounting medium, containing 2.5 μg/mL 4’,6-diamidino-2-phenylindole (DAPI) (Molecular Probes D1306) for nuclear visualisation.

Mitchell R., Mellows B., Sheard J., Antonioli M., Kretz O., Chambers D., Zeuner M.T., Tomkins J.E., Denecke B., Musante L., Joch B., Debacq-Chainiaux F., Holthofer H., Ray S., Huber T.B., Dengjel J., De Coppi P., Widera D, & Patel K. (2019). Secretome of adipose-derived mesenchymal stem cells promotes skeletal muscle regeneration through synergistic action of extracellular vesicle cargo and soluble proteins. Stem Cell Research & Therapy, 10, 116.

+ Open protocol

+ Expand

Protein Expression Profiling of Primed Smooth Muscle Cells

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

Expression of p70S6K, phospho-p70S6K, NFκB-p65 and phospho-NFκB-p65 in primed smooth muscle cells were measured in cell lysate by western blot. Cells were primed with oxLDL as described above and lysed after 4 h. To analyze expression of Hif1α, cells were primed with oxLDL as described above and harvested on day 5. 7.5 to 10% acrylamide gels were loaded with equal amount of protein per lane. Proteins were blotted onto a PVDF membrane and blocked in 5% milk (w/v) in Tris-buffered saline supplemented with Tween 20 (TBS-T). Membranes were incubated in primary antibody [p70S6 Kinase (49D7) Rabbit Antibody, Cell Sigaling, 2708; Phospho-p70S6 Kinase (Thr389) Antibody, Invitrogen 710095; phospho-NFκB p65 (Ser536) Antibody, Cell Signaling 3033; NFκB p65 (F-6), Santa Cruz, Sc-8008; Hif-1α Antibody, BD 610959; Vinculin Antibody (7F9), Santa Cruz, Sc-73614] in TBS-T overnight at 4°C, washed 3 times and incubated in secondary antibody (Goat anti-rb IgG- HRP, Santa Cruz, sc-2004; Goat anti-mouse IgG-HRP Santa Cruz, sc-2005) in TBS-T for 1 h. For development of the membranes Pierce ECL Western Blotting Substrate (ThermoFisher, 32106) was used.

Schnack L., Sohrabi Y., Lagache S.M., Kahles F., Bruemmer D., Waltenberger J, & Findeisen H.M. (2019). Mechanisms of Trained Innate Immunity in oxLDL Primed Human Coronary Smooth Muscle Cells. Frontiers in Immunology, 10, 13.

+ Open protocol

+ Expand

Western Blot Analysis of Protein

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

Protein was run on 10% resolving polyacrylamide gels and transferred to PVDF membrane. Membranes were rinsed with PBS and blocked in 5–10% milk, 1 h at room temperature or overnight at 4 °C. Membranes were then incubated with mouse primary antibody in milk with Tween-20 (Ab Buffer) for 1 h at room temperature or overnight at 4 °C, and washed in PBS with 0.1% Tween-20 (PBST). Membranes were then incubated with anti-mouse secondary antibody in Ab Buffer for 1 h at room temperature, followed by an additional five PBST washes. Primary antibodies were: TWIST1, TWIST 2c1a (Santa Cruz Biotechnology sc-81417) 1:250-1:500; for RELA, NF-κB p65 F-6 (Santa Cruz Biotechnology sc-8008) 1:250-1:500; for GFP, GFP B-2 (Santa Cruz Biotechnology sc-9996) 1:1000; for actin, Sigma Aldrich A1978 or 2066. Secondary antibodies were HRP conjugated anti-mouse and anti-rabbit. Protein was detected using Blue Devil Film (Genesee) and ECL Plus (Thermo Fisher) or digital imaging. Quantitation of digital images was performed using the accompanying software from Syngene (Frederick, MD) or Carestream MI (Woodbridge, CT).

Roberts C.M., Shahin S.A., Loeza J., Dellinger T.H., Williams J.C, & Glackin C.A. (2017). Disruption of TWIST1-RELA binding by mutation and competitive inhibition to validate the TWIST1 WR domain as a therapeutic target. BMC Cancer, 17, 184.

+ Open protocol

+ Expand

Quantifying NF-κB Pathway Activation in HHPC and HHK Cells

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

Total cytoplasmic and nuclear protein expression levels of the cultured HHPC and HHK were determined by western blot analysis, as we previously described [10 (link)]. We used NF-κB (p65) (F-6; Santa Cruz), phospho-NF-κB (p65 S529) (44-711G; Invitrogen^TM, Thermo Fisher Scientific), phospho-IKB-α Ser32/36 (5A5; Cell Signaling), and bcl-2 (C-2; Santa Cruz). We also used β-actin (C4; Santa-Cruz), for cytoplasmic and nuclear extract normalization. Protein levels were quantified by Gel-imaging system (BIO-RAD), in each nuclear and cytoplasmic cellular compartment, and expression levels were estimated by Image Lab 4.1 analysis software 4, BIO-RAD). PARP control was omitted as we focused on NF-κB induced oncogenicity rather than apoptosis.

Sasaki C.T., Toman J, & Vageli D. (2016). The In Vitro Effect of Acidic-Pepsin on Nuclear Factor KappaB Activation and Its Related Oncogenic Effect on Normal Human Hypopharyngeal Cells. PLoS ONE, 11(12), e0168269.

+ Open protocol

+ Expand

Immunofluorescence Staining of PPARγ and NF-κB

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

Cells were rinsed in ice-cold PBS followed by 15 min incubation with 4% paraformaldehyde at room temperature. Cell were then briefly rinsed with PBS, incubated for 5 min with 0.2% Triton-100, and washed 3 times, 5 min every time with PBS. Nonspecific binding sites were blocked with 5% BSA for 1 h. Primary antibodies PPARγ (81B8) Rabbit mAb (Cell Signaling Techn no#5468) and NFκB p65 (F-6) (Santa Cruz Biotechnology, Inc., no SC-8008) were diluted at 1:200, and incubated overnight at 4 °C. Following incubation in cells were washed 3 times, 5 min every time with PBST, and incubated with FITC-coupled secondary antibody (be) and Cy3-labeled Goat Anti-Mouse IgG (H + L) (Beytotime, China, diluted at 1:500) for 1 h at RT. Nuclei staining was performed by incubating the cells with DAPI (Beyotime, China) before mounting the processed coverslips onto ethanol-cleaned glass slides using Dabco mountant. The specimens were viewed with an inverted confocal microscope (Olympus, Japan). Image analysis was done with Image J software.

Feng X., Weng D., Zhou F., Owen Y.D., Qin H., Zhao J., W.e.n.Y.u., Huang Y., Chen J., Fu H., Yang N., Chen D., Li J., Tan R, & Shen P. (2016). Activation of PPARγ by a Natural Flavonoid Modulator, Apigenin Ameliorates Obesity-Related Inflammation Via Regulation of Macrophage Polarization. EBioMedicine, 9, 61-76.

+ 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!