Ab7463

Manufactured by Abcam

Sourced in United Kingdom, United States

Ab7463 is a recombinant monoclonal antibody that recognizes the Histone H3 (tri methyl K4) protein. It is intended for use in Western Blotting and Immunohistochemistry applications.

Automatically generated - may contain errors

Lab products found in correlation

5 protocols using ab7463

Immunohistochemical Analysis of Hydrogels and Decellularized Tissues

Check if the same lab product or an alternative is used in the 5 most similar protocols

Hydrogels and decellularized tissues were fixed in 10% neutral buffered formalin for 1 hr at room temperature, dehydrated, paraffin embedded and cut into 5μm sections. For immunostaining, sections were deparrafinized, rehydrated and subjected to heat mediated antigen retrieval in pH 9.0 Tris-EDTA buffer containing 0.05% v/v Tween 20 for 20 mins using a steam cooker. The following antibodies were used: rabbit polyclonal anti-ki67 (3.5μg/ml; Abcam; ab15580) and rabbit monoclonal (EP1601Y) anti-cytokeratin 5 (1.22μg/ml; Abcam; ab52635), and polyclonal goat anti-rabbit alexafluor 488 conjugated secondary (1μg/ml; ThermoFisher; A32731. Sections were all counterstained with DAPI. All images were taken on a Zeiss Axio Observer Z1 microscope and analyzed using the Zeiss Zen software. Cell counts were performed manually and fluorescence intensity was measured using the Zen software on a minimum of 6 random locations per field of view across a minimum of 4 sections of 3 separate samples. For Laminin 111/112 measurements, hydrogels were formed in a chamber slide, and stained with a rabbit polyclonal anti laminin 111/112 antibody (10μm/ml; Abcam; ab7463) and a goat alexafluor 488 conjugated secondary as described above. Fluorescence intensity was performed as described above across 3 independent samples.

Mollica P.A., Creech E., Reid J.A., Zamponi M., Sullivan S.M., Palmer X.L., Sachs P.C, & Bruno R.D. (2019). 3D bioprinted mammary organoids and tumoroids in human mammary derived ECM hydrogels. Acta biomaterialia, 95, 201-213.

+ Open protocol

+ Expand

Immunofluorescence Staining of Tissue Sections

Check if the same lab product or an alternative is used in the 5 most similar protocols

Gels were fixed in 10% neutral buffered formalin, paraffin-embedded, and sectioned. Sections were prepared for staining by deparaffinizing in a xylene substitute, rehydration, and heat-mediated antigen retrieval using pH 9 tris-EDTA with 0.05% tween 20. Sections were blocked in 10% goat serum and incubated with primary antibodies in a humidified chamber at 4 °C overnight. Secondary antibodies were added for 1 h at room temperature. Sections were counterstained with 4′,6-diamidino-2-phenylindole (DAPI). Antibodies were used at the following concentrations: anti-green fluorescent protein (anti-GFP) rabbit IgG Alexa Fluor 488 conjugated (1:75; Invitrogen A21311, Invitrogen, Carlsbad, CA, USA), rabbit polyclonal antibody to GJB1 [Cx32] (1:25; HPA010663, Sigma-Aldrich, St. Louis, MO, USA), biotinylated rabbit polyclonal antibody to red fluorescent protein (RFP) (1:500; Abcam, Cambridge, UK, ab34771), rabbit anti-laminin 1 + 2 (1:100; Abcam, ab7463), and mouse anti-laminin 5 antibody (1:100; Abcam, ab78286), Alexa Fluor 488 and 568 conjugated goat secondary antibodies (1:1000; Thermo Fisher Scientific) or avidin Alexa Fluor 488 (Thermo Fisher Scientific A21370). All sections were counterstained with DAPI and imaged by using a Zeiss axio-observer Z1 fluorescent microscope.

Reid J.A., Mollica P.M., Bruno R.D, & Sachs P.C. (2018). Consistent and reproducible cultures of large-scale 3D mammary epithelial structures using an accessible bioprinting platform. Breast Cancer Research : BCR, 20, 122.

+ Open protocol

+ Expand

Immunofluorescent Staining of Tumor Sections

Check if the same lab product or an alternative is used in the 5 most similar protocols

The deparaffinization, rehydration, and antigen retrieval of tumor sections were as described in IHC. Slides were washed with PBS before incubation with the first primary antibody, rabbit monoclonal anti‐CD34 (1:1000; ab198395, Abcam), at 4°C overnight. After being washed in PBS, the sections were incubated with mouse monoclonal anti‐GFAP (1:1000; ab10062">http://ab10062, Abcam) or anti‐laminin antibodies (1:1000; ab7463">http://ab7463, Abcam). Next, slides were washed with PBS and incubated with Alexa Fluor® 488‐conjugated goat anti‐rabbit IgG (1:1000; O‐11038; Life Technologies, Carlsbad, CA, USA) and Alexa Fluor® 594‐conjugated goat anti‐mouse IgG (1:1000; AK‐A11005; Life Technologies) for 40 min at room temperature. Nuclear staining was performed with 4′,6‐diamidino‐2‐phenylindole (DAPI; D3571; Life Technologies), and the slides were then mounted with Slow Fade Gold antifade reagent (S36938, Thermo Fisher Scientific).

Mei X., Chen Y., Zhang Q., Chen F., Xi S., Long Y., Zhang J., Cai H., Ke C., Wang J, & Chen Z. (2020). Association between glioblastoma cell‐derived vessels and poor prognosis of the patients. Cancer Communications, 40(5), 211-221.

+ Open protocol

+ Expand

Ascorbic acid-induced extracellular matrix proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse embryonic fibroblasts were cultured in DMEM with 10% fetal bovine serum, 2 mM L-glutamine, 0.2 mM penicillin/streptomycin at 37°C in 5% CO₂. At confluence, cells were serum-deprived and treated with 50 μg.mL⁻¹ of ascorbic acid for 16 hours. Proteins were separated on 4–15% gradient SDS-PAGE gels (Bio-Rad Laboratories Inc., Hercules, CA). Membranes were blocked 16 hours in Tris Buffered Saline/0.1% Tween-20 (TBS-T) with 5% non-fat milk, then incubated 16 hours with primary antibodies: rat COL4A1 (1:200, H11 clone, Shigei Medical Research Institute, Japan^{26 (link)}), tubulin (1:10000, T6557, Sigma-Aldrich, St Louis, MO), laminin (1+2) (1:2000, ab7463, Abcam, Cambridge, UK). After 3 washes in TBS-T, membranes were incubated 1 hour with horseradish peroxidase-conjugated secondary antibodies (1:10000, Jackson ImmunoResearch, West Grove, PA). After 3 washes, SuperSignal West Pico Chemiluminescent substrate (Thermo Scientific, Waltham, MA) was used according to manufacturer’s instructions.

Jeanne M., Jorgensen J, & Gould D.B. (2015). Molecular and Genetic Analysis of Collagen Type IV Mutant Mouse Models of Spontaneous Intracerebral Hemorrhage Identify Mechanisms for Stroke Prevention. Circulation, 131(18), 1555-1565.

+ Open protocol

+ Expand

Scaffold Characterization: DNA and ECM

Check if the same lab product or an alternative is used in the 5 most similar protocols

Before and after freeze-drying, GelMA/ECM scaffolds were washed with PBS, incubated with DAPI at 37 °C for 20 min, and photographed with a fluorescence microscope (Carl Zeiss, Germany). Partial scaffolds were added to the lysate, and the steps of the kit were followed for DNA extraction (Tiangen, China). The concentration of DNA was detected by a Nanodrop 2000 (Thermo Fisher Scientific, USA).
Before and after freeze-drying, GelMA/ECM scaffolds were fixed with 4% paraformaldehyde (PFA) (Solarbio, China) and then incubated with anti-laminin antibody (ab7463, Abcam, USA) or anti-collagenIantibody (ab260043, Abcam, USA) overnight at 4 °C. After removal of excess antibodies, the samples were incubated with secondary antibodies (ab150079, ab150077, Abcam, USA) at 37 °C for 2 h. A fluorescence microscope was used to collect the images.

Chen Z., Wang L., Chen C., Sun J., Luo J., Cui W., Zhu C., Zhou X., Liu X., Yang H., & Shi Q. (2022). NSC-derived extracellular matrix-modified GelMA hydrogel fibrous scaffolds for spinal cord injury repair. NPG Asia Materials, 14(1).

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!