Rabbit anti runx1

Manufactured by Abcam

Rabbit anti-Runx1 is a primary antibody targeting the Runx1 protein. Runx1 is a transcription factor that plays a crucial role in the regulation of hematopoiesis and the development of various blood cell lineages. This antibody can be used in techniques such as Western blotting, immunohistochemistry, and immunocytochemistry to detect and study the Runx1 protein.

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5 protocols using rabbit anti runx1

Immunohistochemical Analysis of Lung Tissue

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Paraffin-embedded lung sections (4-µm thickness) were used for this study. Images were taken under polarized light using an upright dry 40× objective. Tissues were deparaffinized and after antigen retrieval, permeabilized with 0.4% triton X-100. Tissues were blocked with 10% BSA and maintained in PBS + 5% BSA + 0.4% triton X-100 throughout antibody treatments. Primary antibodies were incubated at 4°C overnight and secondary antibodies were incubated for 1 hr at room temperature. Primary antibodies used (1:200 dilution) include rabbit anti-NFκB p65 (#8242, Cells Signaling Technology); mouse anti-cRel (#MA5-15859, Thermo Fisher Scientific); rabbit anti-NFκB1 (#13586, Cells Signaling Technology); mouse anti-NFκB1 (#NBP2-66976, Novus); rabbit anti-RUNX1(#ab229482, Abcam); goat anti-IL33 (#AF3626 R&D); mouse anti-ICAM1 (#sc-1511, Santa Cruz Biotechnology, Inc); and mouse anti-CD3 ((# sc-7296, Santa Cruz Biotechnology, Inc). Goat anti-rabbit and anti-mouse IgG-A594 or IgG-A488 were used as secondary antibodies (1:200 dilution at RT). For anti-goat, rabbit anti-goat IgGA-647 was used as secondary antibody (1:200 dilution at RT). DAPI was used for nuclear counterstaining. The ProLong Gold antifade reagent was used as mounting media. Mount slides were examined with a Leica DM6000 B. Slide Book 6 (3i) was used for analysis and capturing images.

Verma M., Verma D., Sripada A.S., Sirohi K., Varma R., Sahu A, & Alam R. (2023). NFκB1 inhibits memory formation and supports effector function of ILC2s in memory-driven asthma. Frontiers in Immunology, 14, 1217776.

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Immunohistochemistry and Immunofluorescence Assays

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Immunohistochemistry (IHC) and immunofluorescence (IF) were performed as previously described.^{10 (link)} The following primary antibodies were used: rabbit anti-Runx1 (Abcam, ab23980), mouse anti-Col2α1 (Santa Cruz, sc-52658), rabbit anti-MMP13 (Abcam, ab39012), rabbit anti-ADAMTS5 (Santa Cruz, sc-83186), rabbit anti-Yap (Cell Signaling Technology, 14074 S), p-Smad2/3 (Cell Signaling Technology, 8828 S), and mouse anti-active-β-catenin (Millipore, 05–665). The secondary antibodies were goat anti-rabbit IgG-FITC, goat anti-rabbit IgG-TRITC, goat anti-mouse IgG-FITC and goat anti-mouse IgG-TRITC from Santa Cruz. Images were taken by a Leica DMLB microscope and a Leica D3000 fluorescence microscope. ImageJ software was used to perform counts for the quantification of IHC or IF staining.

Zhang Y., Zuo T., McVicar A., Yang H.L., Li Y.P, & Chen W. (2022). Runx1 is a key regulator of articular cartilage homeostasis by orchestrating YAP, TGFβ, and Wnt signaling in articular cartilage formation and osteoarthritis. Bone Research, 10, 63.

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Western Blot Analysis of Chondrocyte Markers

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Proteins were loaded on SDS‒PAGE gels and then electrotransferred to nitrocellulose membranes. We used the following primary antibodies: rabbit anti-Runx1 (Abcam, ab23980), rabbit anti-MMP13 (Abcam, ab39012), mouse anti-Sox9 (Santa Cruz, sc-166505), rabbit anti-Col10ɑ1 (Thermo Fisher, PA5-115039), rabbit anti-Yap (Cell Signaling Technology, 14074 S), and mouse anti-β-Tubulin (Santa Cruz, sc-166729). The secondary antibodies were goat anti-rabbit IgG-HRP (sc-2004) and rabbit anti-mouse IgG-HRP (sc-358917) from Santa Cruz.

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Sensory Neuron Characterization by ISH and IF

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Mice were anesthetized with isoflurane and perfused with 4% paraformaldehyde in PBS (pH 7.4). DRGs were dissected and post-fixed at 4°C for 3–5 h. Tissues were dehydrated in 20% sucrose at 4°C overnight, embedded in O.C.T., and then they were stored at−80°C before use. The detailed procedures for in situ hybridization (ISH) and immunofluorescence (IF) staining have been described previously (Chen et al., 2006 (link)). Probes for TrkA, MrgprA3, MrgprB4, MrgprC11, MrgprD, Nppb, Sst, Cysltr2 and TH were used. The probe templates were amplified from cDNAs prepared from the mouse DRG. The probes were synthesized in vitro with a digoxigenin (Roche) label. The primary antibodies used for IF staining included rabbit anti-Runx1 (1:50, Abcam), rabbit anti-NF200 (1:1000, Sigma), and rabbit anti-CGRP (1:200, Peninsula) antibodies. The secondary antibodies included an Alexa Fluor 488-conjugated goat anti-rabbit antibody (1:1000, Molecular Probes), Alexa Fluor 568-conjugated goat anti-rabbit antibody (1:1000, Molecular Probes), TSA plus Fluorescein Reagent (1:1000, AKOYA), TSA plus Cy3 Reagent (1:1000, AKOYA), and Cyanine5 Amplific action Reagent (1:500, AKOYA). All primary antibodies and secondary antibodies were incubated with sections overnight at 4°C and 1.5 h at RT, respectively. Images were collected using a Zeiss confocal microscope (Zeiss LSM 800).

Cao Z., Huang C., Lu F., Jiang X., Hu Y., Cao C, & Liu Z. (2022). Meis1 Regulates Nociceptor Development and Behavioral Response to Tactile Stimuli. Frontiers in Molecular Neuroscience, 15, 901466.

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Antibodies for Immunoblotting and Immunofluorescence

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The following primary antibodies were used for immunoblotting or immunofluorescence staining: goat anti-Lyve1 (R&D Systems, #AF2125; dilution 1:250), rabbit anti-phospho AKT (Ser 473, Cell Signaling, 4060S; dilution 1:1000), rabbit anti-AKT (Cell Signaling, 4685; dilution 1:1000), rabbit anti-phospho ERK1/2 (Cell Signaling, 4370; dilution 1:1000), rabbit anti-ERK1/2 (Cell Signaling, 4695; dilution 1:1000), rabbit anti-tubulin (Cell Signaling, 2148; dilution 1:2000), rabbit anti-GAPDH (Cell Signaling, 2118; dilution 1:1000), chicken anti-GFP (abcam ab13970; 1:4000), rat anti-endomucin (Santa Cruz, sc-65495; dilution 1:50), rabbit anti-Prox1 (AngioBio, 11-002; dilution 1:1000), rabbit anti-neuropilin-2 (Cell Signaling, 3366; dilution 1:500), rabbit anti-Runx1 (Abcam, ab92336; dilution 1:1000), and rabbit anti-KRAS^G12D (Cell Signaling, #14429; dilution 1:1000).

Fernandes L.M., Tresemer J., Zhang J., Rios J.J., Scallan J.P, & Dellinger M.T. (2023). Hyperactive KRAS/MAPK signaling disrupts normal lymphatic vessel architecture and function. Frontiers in Cell and Developmental Biology, 11, 1276333.

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