The tissue is fixed in formalin and embedded in paraffin, and then cut into thin slices. The sections were deparaffinized with xylene and hydrated with ethanol, and then the antigen was recovered by pressure cooking. At room temperature, the sections were sliced with TNC primary antibody (ab108930, Abcam, Cambridge, MA), MMP9 (Abcam, ab76003, 1:1000 dilution), MMP2 (Abcam, ab92536, 1:1000 dilution) and CD31 (ab134168, Abcam), incubated for 60 min and then incubated with IgG H&L (HRP; 1:200 dilution, Abcam, Cambridge, UK). Then, the sections were stained with chromogen and counterstained with hematoxylin. The score is based on the intensity of staining (0 means no staining, 1 means weak staining, 2 means moderate staining, and 3 means strong staining). The product of the two levels was calculated as the final expression score.
Ab108930
Manufactured by Abcam
Sourced in United Kingdom
Ab108930 is a primary antibody for the detection of a specific protein. It is designed for use in various laboratory techniques, such as immunohistochemistry and Western blotting, to identify and analyze the target protein in samples.
Automatically generated - may contain errors
Lab products found in correlation
Ab76003, by Abcam (4 mentions)
Ab8226, by Abcam (3 mentions)
Ab134168, by Abcam (2 mentions)
Ab92536, by Abcam (2 mentions)
Ab58655, by Abcam (2 mentions)
Ab69506, by Abcam (2 mentions)
Enhanced chemiluminescence reagent, by Fdbio (2 mentions)
G5001, by Wuhan Servicebio Technology (2 mentions)
Ab5694, by Abcam (2 mentions)
Ab34710, by Abcam (2 mentions)
Ab7778, by Abcam (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Ab216347, by Abcam (1 mentions)
Ab27568, by Abcam (1 mentions)
Ab50887, by Abcam (1 mentions)
Chemiluminescence detection reagent, by Merck Group (1 mentions)
Ab18203, by Abcam (1 mentions)
Ab181602, by Abcam (1 mentions)
Ab6721, by Abcam (1 mentions)
Ab1416, by Abcam (1 mentions)
23 protocols using ab108930
1
Immunohistochemical Analysis of Tissue Samples
2
Protein Expression Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell lysates were collected, and 30 mg protein of each sample was separated using 10% SDS-PAGE. The protein was then electrotransferred onto Polyvinylidene Fluoride (PVDF) membrane (Millipore, Boston, MA, USA). The PVDF membrane was sealed in 10% skimmed milk. Then the membrane was incubated with primary antibodies, including anti-TNC (Abcam, ab108930, 1:1000 dilution), anti-GAPDH (Abcam, ab181602, 1:10,000 dilution), and anti-ERK (Abcam, ab184699, 1:1000 dilution)), anti-p-ERK (Abcam, ab201015, 1:1000 dilution), anti-MMP9 (Abcam, ab76003, 1:1000 dilution), anti-MMP2 (Abcam, ab92536, 1:1000 dilution), anti-E-cadherin (Abcam, ab1416, 1:1000 dilution), anti-N-cadherin (Abcam, ab18203, 1:1000 dilution), anti-vimentin (Abcam, ab92547, 1:1000 dilution), anti-snail (Abcam, ab216347, 1:1000 dilution), anti-clumping (Abcam, ab27568, 1:1000 dilution) and anti-TWIST1 (Abcam, ab50887, 1:1000 dilution) overnight. Then the membrane and horseradish peroxidase-conjugated secondary antibody (Abcam AB6721, 1:10,000 dilution) were incubated for 2 h at room temperature. Chemiluminescence detection reagent (Millipore, Boston, MA, USA) was used to visualize protein bands.
3
Proteomic Analysis of GBM Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GBM tissues (n = 4) cryopreserved in liquid nitrogen after surgery at the First Affiliated Hospital of Wenzhou Medical University were collected. Clinical information for 4 patients is presented in Supplementary Table 2 . Total proteins were extracted and quantified using a bicinchoninic acid (BCA) assay. The antibodies used were: anti-TNC (Abcam ab108930), anti-SSTR2 (Abcam ab229007), anti-SERPINA3 (Abcam ab205198), anti-TNFRSF19 (Abcam ab96220), and anti-GAPDH (Abcam ab8245). All primary antibodies were rabbit anti-human antibodies. The secondary antibody was a goat anti-rabbit antibody. Briefly, equal amounts (40 μg) of protein from each sample (tumors and control tissues) were separated by 10% SDS-PAGE electrophoresis and then transferred to PVDF membranes. The PVDF membranes were incubated with the primary antibodies, followed by the secondary antibodies, and then visualized. GAPDH was used as an internal reference for the western blot analysis.
4
Multiplexed Immunofluorescence Staining of Frozen Tissue
5
Immunodetection of Neurological Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used for immunodetection: rabbit antibodies against glial fibrillary acidic protein (GFAP) (1:2500, Sigma-Aldrich, G9269, Saint Louis, USA); rabbit antibodies to ionized calcium binding adaptor molecule 1 (Iba1) (1:500, Wako Chemicals, 019–19,741, USA); rabbit antibodies against Ki67 (1:250, Abcam, ab16667, Cambridge, UK); rabbit antibodies to tenascin-C (1:100, Abcam, ab108930, Cambridge, UK); rabbit antibodies to TMEM119 (1:5000, Proteintech, 66,948–1-Ig, IL, USA); rabbit antibodies to CCL2 (1:500, Novus bio, NBP1-07,035, Milan, Italy); rabbit antibodies against MHC Class II (MHCII) (1:500, Abcam, ab180779, Cambridge, UK); rabbit antibodies to MMP9 (1:500, Invitrogen, JA80-73, Waltham, USA); rabbit antibodies to Fibulin-2 (1:500, Invitrogen, AB_11153545, Waltham, USA); mouse antibodies to β-Actin (1:10,000, Sigma, Saint Louis, USA).
6
Fibroblast Immunofluorescence Analysis of Tenascin-C
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fibroblasts seeded on 8-well Lab-Tek II chamber glass slides (Nalgene Nunc International) were incubated in serum-free DMEM with or without tenascin-C (2 μg/ml) for 72 hours. Cells were then fixed, permeabilized, and incubated with antibodies against ASMA (Abcam, 1:500, ab5694), type I collagen (Southern Biotechnology, 1:100, 1310-01), RP105 (Abcam, 1:100, ab184956), and MD1 (Santa Cruz, 1:50, sc-390613), followed by Alexa Fluor–labeled secondary antibodies (Invitrogen) as described previously (18 (link)). For tissue immunofluorescence analyses, paraffin-embedded skin sections were incubated with ASMA (Abcam, 1:100, ab5694), p-p65 (Cell Signaling Technology, 1:100, 3033), Fn-EDA (Abcam, 1:100, ab6328), tenascin-C (Abcam, 1:100, ab108930), RP105 (Abcam, 1:100, ab184956), and MD1 (Santa Cruz, 1:50, sc-390613), followed by appropriate secondary antibodies. Nuclei were detected using DAPI. Slides were mounted, and immunofluorescence and immunohistochemistry were evaluated under Nikon A1R laser scanning confocal microscope in a blinded manner. Negative controls stained without primary antibodies were used to confirm immunostaining specificity.
7
Western Blot Analysis of TNC in H9C2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The total protein of H9C2 cells were lysed using RIPA Buffer (AR0105) and measured the concentration of lysate by BCA Protein Assay Kit (AR1189A). The protein sample was separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred onto the PVDF membranes (AR0136-02) which were subsequently submerged in Blocking Buffer (AR0174) for 1 h at room temperature, and then were incubated with primary antibodies of TNC (ab108930, 1/1000, 241 kDa, Abcam, Cambridge, UK) and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, ab9484, 1/10000, Abcam, UK) at 4 °C overnight. Afterwards, the membranes were incubated with Goat Anti-Rabbit IgG (ab205718, 1/2000, Abcam, UK) and Goat Anti-Mouse IgG (ab205719, 1/2000, Abcam, UK) at room temperature for 2 h iBright Imaging System (CL750, Invitrogen, USA) and ECL Substrates (AR1191) were used to develop protein bands. GAPDH served as the internal control. All reagent products used were supplied by Boster Biological Technology (China).
8
Immunohistochemical Analysis of Glioma Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IHC staining was performed as reported previously35 (link). The expression level of TNC in glioma tissues was scored as the proportion of the area with positive staining (0–100%) multiplied by the staining intensity (0, negative; 1, weak; 2, moderate; 3, intense). The scores were determined by two pathologists independently. The median score was chosen as the cut-off value for defining high and low expression. Sections were probed with primary anti-TNC (ab108930, Abcam, Cambridge, MA), anti-CD31 (ab134168, Abcam), anti-pAkt (4060P, Cell Signaling Technologies [CST], Danvers, MA,), anti-matrix metalloproteinase (MMP) 2 (ab86607, CST), and anti-MMP9 (ab76003, Abcam) antibodies overnight at 4 °C, and the antibodies were detected using the DAB system (Golden Bridge, Beijing, China).
9
Immunohistochemical Analysis of Tenascin C and Scleraxis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemistry was performed using rabbit anti-human tenascin C monoclonal (ab108930; Abcam, Cambridge, UK) or rabbit anti-human scleraxis polyclonal (ab58655; Abcam) antibodies. Tissue sections on slides were deparaffinized, incubated with methanol and 0.3% hydrogen peroxide (H2O2) for 30 min, and washed. The slides were then incubated with the respective primary antibodies (1:200) at 4 °C overnight. After additional washing steps, the slides were further incubated with horseradish-conjugated secondary polyclonal rabbit immunoglobulin G antibodies (ImmPRESS reagent anti-rabbit Ig kit; Vector Laboratories Burlingame, CA, USA). After additional washing steps, immunoreactivity was visualized using a diaminobenzidine peroxidase substrate kit (Vector Laboratories), followed by counterstaining with hematoxylin. Negative controls were samples immunostained without the primary antibody.
10
Quantitative Western Blot Analysis of Extracellular Matrix Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analysis was performed using a standard protocol36 (link). Briefly, equal amounts of protein (30 μg) were separated via SDS-PAGE and electro-transferred onto polyvinylidene difluoride membranes (EMD Millipore). Membranes were probed with primary anti-TNC (ab108930, Abcam), anti-MMP2 (ab86607, Abcam), anti-MMP9 (ab76003, Abcam), anti-pan-Akt (4691 s, CST), anti-pAkt473 (4060 P, CST), anti-pAkt308 (sc-135650, Santa Cruz Biotechnology, Santa Cruz, CA), anti-β-actin (8457 S, CST) antibodies and then probed sequentially with secondary goat anti-mouse and anti-rabbit antibodies (7076P2, 7074P2, CST) and visualized using an enhanced chemiluminescence kit (Beyotime, Shanghai, China).
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!