A total of 25 μg protein from each sample was separated on 10% Bis-Tris polyacrylamide gel through electrophoresis and then blotted onto polyvinylidene fluoride (PVDF) membranes (GE Healthcare, Piscataway, NJ, USA). Then, the membrane was blocked with 5% (5 g/100 mL) nonfat dry milk (Bio-Rad, CA, USA) in tri-buffered saline plus Tween (TBS-T) buffer for 2 h. Blots were immunostained with primary antibody at 4°C overnight and with secondary antibody at room temperature for 1 h. Immunoblots were visualized using Immobilon™ Western Chemiluminescent HRP Substrate (Millipore). The primary antibodies used for western blotting were rabbit rabbit anti-human Fibronectin antibody (#sc-8422, 1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit anti-human β-actin antibody (#sc-47778, 1:1000; Santa Cruz Biotechnology). Horseradish peroxidase-conjugated (HRP) anti-rabbit antibodies (1:5000; Santa Cruz Biotechnology) were used as the secondary antibodies. Protein levels were normalized to β-actin.
Rabbit anti human β actin antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
The Rabbit anti-human β-actin antibody is a primary antibody that recognizes the β-actin protein, which is a widely expressed cytoskeletal protein found in eukaryotic cells. This antibody can be used to detect and quantify β-actin levels in various sample types, such as cell lysates or tissue extracts, through techniques like Western blotting, immunohistochemistry, or immunocytochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Horseradish peroxidase conjugated hrp anti rabbit antibodies, by Santa Cruz Biotechnology (3 mentions)
Pvdf membrane, by GE Healthcare (2 mentions)
Non fat dry milk, by Bio-Rad (2 mentions)
Immobilon western chemiluminescent hrp substrate, by Merck Group (2 mentions)
Hrp conjugated goat anti rabbit igg secondary antibody, by Santa Cruz Biotechnology (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Image lab 5, by Bio-Rad (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Ripa lysis buffer, by BestBio (1 mentions)
Trans blot cell system, by Bio-Rad (1 mentions)
Ecl reagent, by Thermo Fisher Scientific (1 mentions)
Phosphates inhibitor cocktail, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Rabbit anti human bax antibody, by Cell Signaling Technology (1 mentions)
Rabbit anti human ezh2 antibody, by Cell Signaling Technology (1 mentions)
Bca protein assay kit, by Promega (1 mentions)
Ripa buffer, by Merck Group (1 mentions)
Protease inhibitor, by Merck Group (1 mentions)
Rabbit anti human e cadherin, by Santa Cruz Biotechnology (1 mentions)
Immobilon p membrane, by Thermo Fisher Scientific (1 mentions)
6 protocols using rabbit anti human β actin antibody
1
Western Blot Analysis of Fibronectin
2
Placental AQP3 Protein Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total cytoplasmic protein was extracted from placental tissue using RIPA lysis buffer (BestBio Co., Ltd.; http://www.bestbio.com.cn/ ) and quantified using a BCA Protein Assay kit (Shanghai Guangrui Biotechnology Co., Ltd.). Protein (20 µl/lane) was denatured and resolved by 8% SDS-PAGE (Shanghai Huyu Biotechnology Co., Ltd.; http://www.shhymall.com/. ), and transferred at 240 mA for 2 h onto a PVDF membrane (Bio-Rad Laboratories, Inc.). Non-specific regions of the PVDF membrane were blocked by incubating the membrane with 5% skimmed milk at room temperature for 2 h. Then, the PVDF membrane was probed with primary rabbit anti-human AQP3 antibody (cat. no. sc518001; Santa Cruz Biotechnology Inc.; 1:1,000) or rabbit anti-human β-actin antibody (cat. no. sc58675; Santa Cruz Biotechnology Inc.; 1:5,000) overnight at 4°C, followed by incubation with goat anti-rabbit IgG HRP-conjugated secondary antibodies (cat. no. 111-035-003; Jackson ImmunoResearch Laboratories, Inc.; 1:2,000) for 2 h at room temperature. Finally, detection of protein signals was performed using an ECL Chemiluminescence kit (GrBio), and observed by Image Lab5.0 software (Bio-Rad Laboratories, Inc.). To obtain the relative expression of AQP3, the AQP3 signal was normalized to the β-actin signal.
3
Western Blot Analysis of TOM20 in Sorted GlycoA+ NRBC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The proteins of sorted GlycoA+ NRBC were extracted by lysis buffer (Biotech, Beijing, China). Protein concentration was determined using the Pierce BCA Protein Assay Kit (Thermo Scientific, USA). Samples were separated by 12% SDS-PAGE gels and then transferred to PVDF membranes (PE, USA) by a Trans-Blot Cell system (Bio-Rad, USA) using standard Western blotting procedures. The membranes were probed with a rabbit anti-human TOM20 antibody (Cell Signaling Technology, number 13929, USA) at 1 : 1000 and incubated overnight at 4°C. The rabbit anti-human β-actin antibody (Santa Cruz Biotechnology Inc., sc-47778, USA) at 1 : 5000 was used as loading control. The goat anti-rabbit IgG HRP-conjugated secondary antibody (Santa Cruz Biotechnology Inc., sc-2054, USA) was incubated for 1 hour at room temperature. After washing, an electrochemiluminescence (ECL) reagent (Thermo Scientific, number 32106, USA) was used for chemiluminescence.
4
FOXO3 Expression in NSLCL Cells Treated with ESMC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NSLCL cells treated with ESMC for 48 h were lysed with RIPA lysis buffer containing protease inhibitor cocktail and phosphates inhibitor cocktail (Sigma-Aldrich; Merck KGaA) on ice for 30 min, then lysis buffer was collected, and centrifuged at 12,000 g, 4°C for 10 min. The protein lysates were resolved by SDS-PAGE, and separated proteins were transferred to PVDF membranes and blocked with 5% skimmed milk for 2 h. The primary antibodies used for western blotting were rabbit anti-human FOXO3 antibody (1:1,000; Santa Cruz Biotechnology, Inc.) and rabbit anti-human β-actin antibody (1:1,000; Santa Cruz Biotechnology, Inc.). Horseradish peroxidase-conjugated (HRP) anti-rabbit antibodies (1:5,000; Santa Cruz Biotechnology, Inc.) were used as the secondary antibodies. The blots were incubated with the respective antibodies overnight at 4°C under gently shaking. Finally, the proteins were detected by using horseradish peroxidase labeled secondary antibodies and an enhanced chemiluminescence detection system.
5
Western Blot Analysis of EZH2 and BAX
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LAD cells were lysed with radioimmunoprecipitation assay (RIPA) buffer (Sigma-Aldrich) containing protease inhibitors (Sigma-Aldrich). Protein quantification was done using a BCA protein assay kit (Promega). A total of 25 μg protein from each sample was separated on 10% Bis-Tris polyacrylamide gel through electrophoresis and then blotted onto polyvinylidene fluoride (PVDF) membranes (GE Healthcare, Piscataway, NJ, USA). Then, the membrane was blocked with 5% (5 g/100 mL) nonfat dry milk (Bio-Rad, CA, USA) in tri-buffered saline plus Tween (TBS-T) buffer for 2 h. Blots were immunostained with primary antibody at 4 °C overnight and with secondary antibody at room temperature for 1 h. Immunoblots were visualized using ImmobilonTM Western Chemiluminescent HRP Substrate (Millipore). Protein levels were normalized to β-actin. The primary antibodies used for western blotting were rabbit anti-human EZH2 antibody (#4905S; 1:1000; Cell Signaling Technology), rabbit anti-human BAX antibody (#2772; 1:1000, Cell Signaling Technology), and rabbit anti-human β-actin antibody (#sc-47778; 1:1000; Santa Cruz Biotechnology, Santa Cruz, CA, USA). Horseradish peroxidase-conjugated (HRP) anti-rabbit antibodies (1:5000; Santa Cruz Biotechnology) were used as the secondary antibodies.
6
Western Blot Analysis of EGFR and E-cadherin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed in radioimmunoprecipitation assay buffer. Cellular proteins were collected and subjected to 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and then electrotransferred onto Immobilon-P membranes (Invitrogen; Thermo Fisher Scientific, Inc.). The membranes were then incubated for 2 h at 37°C with rabbit anti-human EGFR (catalog no., sc-367974; 1:1,000 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) or rabbit anti-human E-cadherin (catalog no., sc-7870; 1:1,000 dilution; Santa Cruz Biotechnology, Inc.). This was followed by incubation with horseradish peroxidase-conjugated donkey anti-rabbit immunoglobulin G secondary antibody for 1 h at 37°C (catalog no., NA934; 1:1,000 dilution; Amersham Pharmacia Biotech, Piscataway, NJ, USA). The rabbit anti-human β-actin antibody (catalog no., sc-7210; 1:1,000 dilution; Santa Cruz Biotechnology, Inc.) was used as an internal marker for control purposes.
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!