Immunochemistry was performed using specific, non-cross reacting rabbit polyclonal antibody against SMMHC (ab124679, Abcam, MA, 1:100 dilution). Appropriate positive controls for SMMHC (human colon) and negative controls (rabbit IgG from DAKO, CA) were included. Sections were examined under an Olympus BX 50 light microscope (Olympus America, Center Valley, PA), and the expression of SMMHC was quantified in 20 random high-power fields (HPFs) using percentage of positive cells stained per HPF. Using the intensity of cells immunostained with SMMHC, a semi quantitative score was used to grade as follows; grade 0: absent, grade 1: 25% stained, grade 2: 26%–50% stained, grade 3: >50 stained.
Ab124679
Manufactured by Abcam
Sourced in United Kingdom
Ab124679 is a rabbit monoclonal antibody that recognizes a specific target. This antibody is designed for use in various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Bx50 light microscope, by Olympus (1 mentions)
Anti bax, by Cell Signaling Technology (1 mentions)
Ge image quant las4000 mini system, by GE Healthcare (1 mentions)
Nbp1 80306, by Novus Biologicals (1 mentions)
Nb100 92142, by Novus Biologicals (1 mentions)
Anti β tubulin, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Ab5694, by Abcam (1 mentions)
Ab124762, by Abcam (1 mentions)
Ab53115, by Abcam (1 mentions)
Ab66133, by Abcam (1 mentions)
Biotin conjugated secondary antibody, by Vector Laboratories (1 mentions)
Ab15580, by Abcam (1 mentions)
Abc kit, by Vector Laboratories (1 mentions)
Ab259341, by Abcam (1 mentions)
Cxcl2, by Thermo Fisher Scientific (1 mentions)
Col1a2, by Affinity Biosciences (1 mentions)
Myh11, by Abcam (1 mentions)
4 protocols using ab124679
1
Quantifying Smooth Muscle Myosin Expression
2
Western Blot Analysis of Cell Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western Blot Analysis was performed to detect SM α-actin, SM-MHC, C-Myb, BCL-2, Bax, β-tubulin, and β-actin protein levels. In brief, total proteins were extracted from cultured cells and measured using a BCA Protein Assay kit (Thermo, USA). Equal amounts of protein were subjected to 6% sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Bedford, MA, USA). The membranes were blocked with 5% BSA for 1 h, followed by incubation with primary antibodies overnight at 4°C. The membranes were incubated with HRP-conjugated secondary antibody (A00098, GenScript, USA). The bands were imaged and analyzed on a GE Image-Quant LAS4000 mini system (GE Healthcare, USA). The primary antibodies used in the study were displayed as follows: anti-SM α-actin (ab5694, Abcam, UK), anti-SM-MHC (ab124679, Abcam, UK), anti-C-Myb (NBP1-80306, Novus, USA), anti-BCL-2 (NB100-92142, Novus, USA), anti-Bax (#2772, Cell Signaling Technology, USA), anti-β-tubulin (#2128, Cell Signaling Technology, USA), and anti-β-actin (#4970, Cell Signaling Technology, USA).
3
Immunohistochemical Characterization of Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh sections (formalin-fixed, paraffin embedded) were dewaxed following standard protocols and antigen retrieval with pH6 citrate retrieval buffer was undertaken for SMMHC, CK14, p63 and Ki67 staining. Sections were stained with anti-α-smooth muscle actin (1 μg/ml, Abcam, ab66133), anti-smooth muscle myosin heavy chain 1+2 (1 μg/ml, Abcam, ab124679), anti-Ki67 (1 μg/ml, Abcam, ab15580), anti-cytokeratin 14 (5 μg/ml, Abcam, ab53115), anti-p63 (0.75 μg/ml, Abcam, ab124762), or with isotype control antibodies, overnight at 4°C. Biotin-conjugated secondary antibodies (Vector Laboratories, CA, USA) were applied for 1 h and signal was amplified by conjugation of HRP (ABC kit, Vector Laboratories). Antibodies were visualized with DAB prior to nuclear counterstaining with hematoxylin.
4
Western Blot Analysis of SMC Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For Western blot analysis, proteins of SMCs were separated by 8% SDS-polyacrylamide gel electrophoresis and then transferred onto polyvinylidene fluoride (PVDF) membranes of 0.45 μm pore size. The membranes were blocked by 5% non-fat milk in TBS-T (20 mM Tris, 137 mM NaCl and 0.1% Tween-20) for 2 h and then incubated with primary antibodies overnight at 4 °C. Followed by TBS-T washes and incubation with horseradish peroxidase (HRP)-conjugated secondary antibodies (Beyotime, China) for 1 h. The immunoblot signal was visualized using a chemiluminescence reagent (Pierce, USA) and ChemiScope 6100 imaging system (Qinxiang Instruments, Shanghai, China).
The following antibodies were used: MYH11 (ab124679, Abcam, Cambridge, UK), α-SMA (#48938, CST, Danvers, MA, USA), CNN1 (24855-1-AP, Proteintech, Rosemont, IL, USA), COL1A2 (AF7001, Affinity Biosciences, Cincinnati, OH, USA), CXCL2 (PA5-79109, Thermo, Carlsbad, CA, USA), IL-6 (ab259341, Abcam, Cambridge, UK) and β-actin (66009, Proteintech, Rosemont, IL, USA).
The following antibodies were used: MYH11 (ab124679, Abcam, Cambridge, UK), α-SMA (#48938, CST, Danvers, MA, USA), CNN1 (24855-1-AP, Proteintech, Rosemont, IL, USA), COL1A2 (AF7001, Affinity Biosciences, Cincinnati, OH, USA), CXCL2 (PA5-79109, Thermo, Carlsbad, CA, USA), IL-6 (ab259341, Abcam, Cambridge, UK) and β-actin (66009, Proteintech, Rosemont, IL, USA).
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!