Spermidine (S0266), 3‐methyladenine (M9281), TBHP (458139) and type II collagenases (1148090) were purchased from Sigma‐Aldrich (St Louis, MO, USA). p62 (ab56416), LC‐3(ab128025), Beclin‐1(ab6242), Atg7(ab52472), cleaved caspase‐3(ab32042), Bax(ab32503) and Bcl‐2(ab59348) antibodies were purchased from Abcam (Cambridge, UK). FITC‐labelled and horseradish peroxidase‐labelled secondary antibodies were purchased from Abcam (ab7086, ab191866, ab193651). 4′, 6‐diamidino‐2‐phenylindole (DAPI) was obtained from Beyotime (C1002, Shanghai, China).
Ab7086
Manufactured by Abcam
Sourced in United Kingdom
Ab7086 is a mouse monoclonal antibody that recognizes the human CD4 antigen. CD4 is a glycoprotein expressed on the surface of T helper cells, monocytes, and macrophages. This antibody can be used for the detection of CD4 in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab25022, by Abcam (2 mentions)
Ab13840, by Abcam (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Ab191866, by Abcam (1 mentions)
Ab193651, by Abcam (1 mentions)
Bcl 2 ab59348, by Abcam (1 mentions)
Fv1000, by Olympus (1 mentions)
Fluorescent microscope, by Leica (1 mentions)
Hoechst 33258, by Merck Group (1 mentions)
Ab124964, by Abcam (1 mentions)
Ab285390, by Abcam (1 mentions)
Ab32575, by Abcam (1 mentions)
Ab6786, by Abcam (1 mentions)
Ab119348, by Abcam (1 mentions)
Ab137855, by Abcam (1 mentions)
Jsm 6380lv, by JEOL (1 mentions)
Triton x 100, by Merck Group (1 mentions)
8 protocols using ab7086
1
Autophagy and Apoptosis Modulation
2
Immunofluorescence Staining of Cultured Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cultured cells were gently washed with cold phosphate-buffered saline (PBS) to remove nonadherent cells before fixation in 2.0% paraformaldehyde for 15 min at room temperature. Then cells were permeabilized in 0.025% Triton X-100 in PBS for 10 min before incubation for 1 h with normal goat serum. After the introduction of diluted primary antibody to the cells overnight at 2–8 °C, the cells were incubated with secondary antibody fluorescein isothiocyanate (Abcam, ab7086, USA) diluted in blocking solution in a dark humidity chamber for 1 h. The sample was counterstained with 4,6-diamidino-2-phenylindole at room temperature for 10 min, and the slides were stored in the dark at 2–8 °C. The micrographs were obtained by laser scanning confocal microscopy (Olympus FV1000, Tokyo, Japan).
3
Immunofluorescent Analysis of PI3K and AKT
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After being fixed with 4% paraformaldehyde for 10 min and permeabilized by 0.3% Triton X-100 for 15 min, MAD-MB-231 and MCF7 cells cultured on glass coverslips were incubated with 3% BSA for 30 min at room temperature to block nonspecific binding. Following that, cells were stained with p-PI3K and p-AKT antibodies overnight at 4°C and fluorescein isothiocyanate- (FITC-) labeled secondary antibodies (1 : 500, ab7086, Abcam) for 2 h at room temperature. Then, the nucleus was counterstained with Hoechst 33258 (Sigma, USA) for 30 min. Cells were photographed under a fluorescent microscope (Leica, Germany).
4
Immunofluorescence Quantification of α-SMA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MLE-12 cells were fixed with 4 % paraformaldehyde (PN4204, G-CLONE) and exposed to BSA (ab245930, Abcam) for 25 min. Then, cells were incubated with antibodies against α-SMA (ab124964, 1/300, Abcam) overnight and FITC-labelled secondary antibody (ab7086, 1/2000, Abcam) for 1 h. Nuclei were dyed with DAPI (ab285390, Abcam), and images were obtained using a microscope (Olympus, Japan). α-SMA expression was quantified using Image J software (National Institutes of Health, Maryland, USA). The formula used was as follows: Mean fluorescence intensity = Integrated Density/Area.
5
Cardiac Fibroblast Immunostaining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After treatment, cardiac fibroblasts were seeded in cell culture dishes, fixed in 4% paraformaldehyde, and permeabilized with 0.1% Triton X-100, followed by blocking in 3% bovine serum albumin (BSA), washed by phosphate-buffered saline (PBS). They were then immunolabeled with specific primary antibodies [alpha smooth muscle actin (α-SMA); 1:200; product code ab32575; Abcam] overnight at 4°C. After rinsing, the cells were incubated with the corresponding FITC-conjugated secondary antibodies (1:250) (product code ab7086; Abcam) in 1% BSA for 1 h at 37°C. Nuclei were stained with DAPI for 5 min at room temperature. Fluorescence was detected under a fluorescence microscope.
6
Cell Viability and Apoptosis Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To investigate cell viability and apoptosis, cells cultured on the scaffold were observed by double staining with fluorescein diacetate (FDA) and propidium iodide (PI). A scanning electron microscope (SEM, JSM-6380LV, JEOL, Tokyo, Japan) was used to visualize the surface features of the scaffold and cells cultured in the scaffold. The samples were visualized with the SEM using an accelerating voltage of 30 kv. For detailed information on SEM sample preparation, see S1 Text . For Immunocytochemistry, the cultures were prepared as previously described [27 (link)]. The primary antibodies used were as follows: polyclonal rabbit anti-cytokeratin 8 antibody (CK8, 1:100, ab137855, Abcam) and monoclonal rat anti-CD44 antibody (1:100, ab119348, Abcam). Then, followed by secondary goat anti-mouse IgG-TRITC (1:1000, ab6786, Abcam), or a goat anti-rabbit IgG-FITC (1:1000, ab7086, Abcam) antibody.
7
Characterization of MVH and c-KIT Expression in Differentiating Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The phenotype of the cells for MVH and c-KIT at days 3 and 7 post-differentiation induction was studied by flow cytometry. For this purpose, at each incubation times of 3 and 7, the cells were harvested by 0.25% trypsin/0.2% EDTA, and incubated with primary anti-mouse MVH (1:200, ab13840, Abcam, Cambridge, UK) and primary anti-mouse c-KIT (1:300, ab25022, Abcam, Cambridge, UK) for 4 hrs at room temperature. The cells were then washed with ice-cold PBS and treated with FITC-conjugated anti-rabbit polyclonal secondary antibody (1:500; ab7086, Abcam, Cambridge, UK) for 2 hrs at room temperature in a dark chamber. The percentage of the cells expressing MVH and c-KIT was determined by flow cytometry. To avoid the interfering effects of the cell population and number difference, the flowcytometry data of the Scaffold/EB and Scaffold/EB/Sertoli were compared with those phenotypic results obtained from groups EB and EB/Ser, respectively. The experiment was conducted in triplicate.
8
Immunocytochemical Quantification of c-KIT and MVH
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The percentage of the cells expressing c-KIT and MVH was determined by ICC and compared among the experimental groups. ICC was performed by a procedure described elsewhere, with a minor modification 13 . In brief, the samples were washed with PBS and fixed with 4% paraformaldehyde (PFA). After permeabilization by 0.1% Triton-X100 (Sigma-Aldrich, St Louis, MO, USA), the samples were treated with 10% goat serum to block non-specific binding sites.
The samples were treated with primary anti-mouse MVH (1:200, ab13840, Abcam, Cambridge, UK) and primary anti-mouse-c-KIT (1:500, ab25022, Abcam, Cambridge, UK) at 4 ⁰C for 24 hrs, and then washed and incubated with fluorescein isothiocyanate (FITC)-conjugated anti-rabbit polyclonal secondary antibody (1:500; ab7086, Abcam, Cambridge, UK) for 2 hrs in the dark. 4′6diamidino-2-phenylindole (DAPI) was used to counterstain the cell nuclei. The ICC slides were viewed under a fluorescent microscope (Olympus BX51, Tokyo, Japan) and the percentage of positive cells for c-KIT and MVH were reported as average cell number per ten high-power field (HPF).
The samples were treated with primary anti-mouse MVH (1:200, ab13840, Abcam, Cambridge, UK) and primary anti-mouse-c-KIT (1:500, ab25022, Abcam, Cambridge, UK) at 4 ⁰C for 24 hrs, and then washed and incubated with fluorescein isothiocyanate (FITC)-conjugated anti-rabbit polyclonal secondary antibody (1:500; ab7086, Abcam, Cambridge, UK) for 2 hrs in the dark. 4′6diamidino-2-phenylindole (DAPI) was used to counterstain the cell nuclei. The ICC slides were viewed under a fluorescent microscope (Olympus BX51, Tokyo, Japan) and the percentage of positive cells for c-KIT and MVH were reported as average cell number per ten high-power field (HPF).
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!