The placental invasion ability assessment was conducted following the methodology outlined in a previous study [18 (link)]. The percentage of interstitial trophoblast invasion into the mesometrial triangle (MT) was utilized to quantify the invasion ability, with infiltrated trophoblast cells identified using a cytokeratin-7 (CK-7) antibody (Abcam, ab181598). Evidence of spiral artery (SA) remodeling was also identified through the presence of CK-7-positive cells arranged on a fibrinoid layer, alongside the absence of α-actin-positive smooth muscle cells. Additionally, the cross-sectional area, as reported by Cotechini et al., was included as an informative indicator [19 (link)]. Both the ratio of the cytokeratin-7-positive trophoblast cell area to the MT area and the cross-sectional areas were measured using image J analysis software.
Ab181598
Manufactured by Abcam
Sourced in United Kingdom, United States
Ab181598 is a recombinant rabbit monoclonal antibody that recognizes Myelin Basic Protein (MBP). It is intended for use in immunohistochemistry (IHC) and Western blotting applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab92547, by Abcam (4 mentions)
Ab16667, by Abcam (3 mentions)
Ab52625, by Abcam (2 mentions)
Ab81289, by Abcam (2 mentions)
Ab150077, by Abcam (2 mentions)
Ab10558, by Abcam (2 mentions)
Ab5694, by Abcam (2 mentions)
Bx51 microscope, by Olympus (2 mentions)
Ab8978, by Abcam (2 mentions)
Triton x 100, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Biosharp (1 mentions)
Bovine serum albumin (bsa), by Boster Bio (1 mentions)
Ab134114, by Abcam (1 mentions)
Ab133582, by Abcam (1 mentions)
Ab134179, by Abcam (1 mentions)
Ab231774, by Abcam (1 mentions)
Ab189524, by Abcam (1 mentions)
Ab40763, by Abcam (1 mentions)
Matrigel, by BD (1 mentions)
27 protocols using ab181598
1
Placental Invasion Ability Assessment
2
Characterization of hAMSCs by Immunofluorescence
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
hAMSCs were fixed with 4% paraformaldehyde (Biosharp, China) for 20 min, and with permeabilized 0.1% Triton-X 100 (Sigma) for 15 min and were blocked with 5% bovine serum albumin (BSA) (Bosterbio) for 30 min at room temperature. The cells were incubated overnight at 4 °C with the primary antibodies anti-Integrin beta 1 (CD29) (ab134179, Abcam), anti-CD44 (ab189524, Abcam), anti-CD73 (ab133582, Abcam), anti-CD105 (ab231774, Abcam), anti-CD19 (ab134114, Abcam), anti-CD34 (ab81289, Abcam), anti-CD45 (ab40763, Abcam), anti-cytokeratin 7 (CK7) (ab181598, Abcam) and anti-cytokeratin 19 (CK19) (ab52625, Abcam) and then incubated with the Alexa Fluor 488-conjugated secondary antibody (ab150077, Abcam) for 1 h at 37 °C in the dark. The cell nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (Sigma) for 5 min. The results were observed by fluorescence microscope.
3
Xenograft Tumor Establishment and Metastasis Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In all the xenograft experiments described, 5 × 106 PBS‐washed cells were resuspended in 200 μl of growth factor reduced Matrigel (BD Biosciences, Allschwil, Switzerland; cat. no. 354230) at the final concentration of 6.5–7.0 mg/ml (depending on the specific lot used) and injected subcutaneously into one flank of 6‐week‐old female mice of the indicated strain. Except where indicated, five mice/condition were used. Mice were purchased from Charles River Laboratories (Saint‐Germain‐sur‐l'Arbresle, France) and maintained in a specific pathogen free area. Experiments were performed according to Institutional ethics guidelines. For the NOD SCID gamma (NSG) experiment, the indicated organs were embedded in paraffin using standard procedures. Two hundreds 5 μm‐thick consecutive sections were cut. Of these, one in every ten was stained with Hematoxylin‐eosin (HE). For the Swiss nude experiment, the procedure was the same, except that 100 5 μm‐thick consecutive sections were cut, and one every ten was stained with HE. HE stained sections were screened for metastasis with the help of an expert pathologist. Immunohistochemistry for cytokeratin 7 or 19 used antibodies ab181598 or ab133496, respectively (both from Abcam, Cambridge, UK), and was performed according to manufacturer's instructions.
4
Quantification of Hepatic Macrophages in Formalin-Fixed Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formalin fixed slides were analyzed for hepatic macrophages using antibodies directed against F4/80 (1:250, T-2028, Rat anti-mouse monoclonal, BMA Biomedicals, Rheinstrasse Switzerland), CK7 (1:500, ab181598, Rabbit monoclonal, Abcam, Boston, MA), and Ki67 (1:100, M7249 TEC3, Rat anti-mouse monoclonal, Dako, Carpinteria, CA) as previously described [36 (link)]. Heat induced antigen retrieval was performed in Dako target Antigen Retrieval Solution (Dako, Carpinteria, CA). Following incubation with primary antibodies overnight, slides were washed 3X5min in tris-buffered saline 1% tween and incubated in HRP-conjugated a goat anti-rat, goat anti-rabbit (MP-7444, MP7401, Vector Laboratories, Newark, CA) secondary antibody for 30 minutes. The peroxidase substrate used was IMMPACT-DAB (SK-4105, Vector Laboratories). Histologic images were captured on an Olympus BX51 microscope equipped with a four-megapixel Macrofire digital camera (Optronics; Goleta, CA) using the Picture Frame Application 2.3 (Optronics). All images were cropped and assembled using Adobe Photoshop Elements (Adobe Systems, Inc.; Mountain View, CA). For quantification of IHC staining, 10 images per animal were obtained at 100X magnification (tiling). All images were imported into SlideBook 6.0 (Intelligent Imaging Innovations, Denver, Colorado; Colin Monks) and pixels per image quantified.
5
Immunohistochemical Analysis of Tissue Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunohistochemistry, paraffin blocks were cut into 4 μm sections, deparaffinized in xylene, and rehydrated in graded ethanol solutions. Endogenous peroxidase activity was blocked with 3% hydrogen peroxide for 20 min, washed with phosphate‐buffered saline (PBS; 0.01 m , pH 7.4) three times for 5 min, and incubated with blocking buffer at 37 °C for 60 min. Tissue sections were incubated overnight at 4 °C with anti‐Desmin (1 : 200, ab15200; Abcam, Guangdong, China), anti‐α‐SMA (1 : 200, ab7817; Abcam), anti‐CK7 (1 : 8000, ab181598; Abcam), anti‐CK19 (1 : 200, ab7755, Abcam), or anti‐EpCAM (1 : 200, ab71916; Abcam) antibodies, and then with horseradish peroxidase (HRP)‐conjugated rabbit anti‐mouse IgG (A9044; 1 : 200) or anti‐rabbit IgG (A0545; 1 : 200) secondary antibodies for 1 h at 37 °C. HRP‐conjugated secondary binding was visualized using a DAB+ substrate chromogen system (Dako, Beijing, China). Nuclei were counterstained with hematoxylin. Bright‐field images were captured using an optical microscope (BX53; Olympus, Shanghai, China). Brown staining was considered positive. Positive areas were analyzed by using winroof software (V6.3, Mitani Corporation, Tokyo, Japan). The positive area (%) per high‐power field (200×) was calculated as follows: expression = positive area/total area × 100%.
6
Placental Histological Analysis in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse placentas were harvested and fixed in 4% paraformaldehyde. Subsequently, they were rinsed in PBS, transferred in ethanol (70%), and processed into paraffin-embedded tissue blocks. Tissue sections of placenta (4 μm) were selected for hematoxylin and eosin (H&E) staining and analyzed by microscopy. Structural analysis of the labyrinth was performed as previously described (55 (link)). Immunohistochemistry was performed using primary antibodies with α-SMA, cytokeratin-7 (ab124964, dilution 1:100; ab181598, dilution 1:8000; Abcam, Cambridge, UK) according to the manufacturer’s protocol. Briefly, sections were deparaffinized and rehydrated. After antigen retrieval, the sections were blocked with 1% bovine serum albumin at room temperature for 1 h. Subsequently, they were incubated with the antibodies at 4°C overnight. The sections were then incubated with a secondary antibody and treated with 3,3′-diaminobenzidine solution for microscopic imaging.
7
Immunohistochemical and Immunofluorescence Analysis of SIRT1 in Placental Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human placental sections were deparaffinized and treated with 3% H2O2 to block the endogenous peroxidase activity. Then, the sections were pretreated heat-mediated antigen retrieval using sodium citrate buffer or Tris/EDTA buffer and incubated with primary antibodies against SIRT1 (dilution 1:100, ab32441, Abcam, UK) at 4 °C overnight. Positive signals were visualized by incubation with HRP-conjugated secondary antibodies using microscope (Olympus IX73).
The mice placental sections were used for immunofluorescence, and the process is similar to immunohistochemistry. The sections were incubated with primary antibodies against CK7 (dilution 1:5000, ab181598, Abcam), SIRT1 (dilution 1:100, ab32441, Abcam) at 4 °C overnight. Positive signals were visualized by incubation with fluorophore-conjugated secondary antibodies. The slides were mounted with mounting medium containing DAPI (Abcam) and then viewed and imaged under a fluorescence microscope (Olympus IX73).
The mice placental sections were used for immunofluorescence, and the process is similar to immunohistochemistry. The sections were incubated with primary antibodies against CK7 (dilution 1:5000, ab181598, Abcam), SIRT1 (dilution 1:100, ab32441, Abcam) at 4 °C overnight. Positive signals were visualized by incubation with fluorophore-conjugated secondary antibodies. The slides were mounted with mounting medium containing DAPI (Abcam) and then viewed and imaged under a fluorescence microscope (Olympus IX73).
8
Immunofluorescence Staining of Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fixed pESCs were incubated with anti-vimentin (1:200, ab92547) and anti-CK7 (1:200, ab181598) (both from Abcam, Cambridge, MA, USA), and fixed ESCs from different treatments were incubated with anti-PRL (1:150, ab64377; Abcam) primary antibody overnight at 4°C separately, washed with PBS and incubated with FITC-conjugated secondary antibody (1:200, ab6717; Abcam). After washing with PBS, cells were mounted with anti-fading mounting medium with DAPI and observed with a fluorescence microscope (magnification, ×40). Images were captured with a charge coupled device camera.
9
Immunofluorescence Staining Protocol for HMGB1 and Cytokeratin 7
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were plated in 6-well plates, each containing 4 sterile 13-mm glass coverslips. When 80% confluent, cells were fixed in 4% paraformaldehyde in PBS for 15 min at RT. Cells were washed 3 times with PBS (137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, and 2 mM KH2PO4) and finally treated with 0.1% Triton/PBS for 15 min at RT. They were then blocked in 1% bovine serum albumin (BSA) in PBS for 1 h at RT. Primary antibodies, anti-HMGB1 (sc-74085; Santa Cruz Biotechnology) or anti-Cytokeratin 7 (ab181598; Abcam, Cambridge, UK) were diluted in 1% BSA in PBS. Cells were incubated with the corresponding primary antibodies overnight at 4 °C, followed by 3 washes with PBS and staining with the secondary antibodies, modified with Alexa Fluor 488 and 568 (Invitrogen, Carlsbad, CA, USA) previously diluted in 1% BSA in PBS for 1 h at RT in the dark. For nuclear staining, after secondary antibody incubation, wells were washed 3 times and stained with Hoechst (Life Technologies, Carlsbad, CA, USA) for 5 min at RT in the dark. Cells were washed once with PBS and once with sterile distilled water. Each coverslip was mounted on a clean slide using ProLong™ Gold Antifade Mountant (Invitrogen). After drying, the slides were stored at 4 °C in the dark until they were examined by confocal microscopy (Nikon A1R). Meander’s correlation coefficient was calculated using Nis Elements software from Nikon.
10
Immunohistochemical Analysis of Endometrial Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human endometrial tissues were fixed with formalin, embedded in paraffin, and sliced into 5-μm sections. Immunohistochemical or immunofluorescence approaches were conducted as previously described [15 (link)]. Briefly, the primary antibodies used are as follows: mouse anti-S100P monoclonal antibodies (MAB2957, R&D Systems, Minneapolis, MN, USA, 1:50 dilution), rabbit anti-cytokeratin monoclonal antibodies (ab181598, Abcam, UK, 1:100), rabbit anti-Vimentin monoclonal antibodies (ab92547, Abcam, UK, 1:100) and isotype antibody as negative control. Slides were then stained with secondary antibody (Cy3-conjugated anti-Mouse IgG, FITC-conjugated anti-Rabbit IgG or HRP-conjugated), followed by DAPI staining or hematoxylin counterstaining for immunofluorescence and immunohistochemistry, respectively. Slide images were visualized using a laser confocal microscope (Leica SP8, Germany) or an upright microscope (Nikon Inc., Japan). Densitometric analysis was conducted to compare the expression level of proteins using Image J (Version 1.5.1; NIH, Bethesda, MD, 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!