Ab104430

Manufactured by Abcam

Ab104430 is a primary antibody product from Abcam. It is a polyclonal antibody raised in rabbit against a synthetic peptide derived from human Sonic Hedgehog. This antibody is intended for use in various research applications.

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Lab products found in correlation

Lsm 710, by Zeiss (1 mentions) Abn78, by Merck Group (1 mentions) Nanozoomer 2.0 ht whole slide scanner, by Hamamatsu Photonics (1 mentions) M9942, by Merck Group (1 mentions) Ab78078, by Abcam (1 mentions) B5002, by Merck Group (1 mentions) Ab16669, by Abcam (1 mentions) Ab13970, by Abcam (1 mentions) Vectashield hardset, by Vector Laboratories (1 mentions) Ab79351, by Abcam (1 mentions) Cfi apo lwd lambda s 40 objective, by Nikon (1 mentions) Af6166, by Abcam (1 mentions) Ab109406, by Abcam (1 mentions) Cm3050s cryostat, by Leica (1 mentions) Superfrost slide, by Avantor (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Optimal cutting temperature compound, by Thermo Fisher Scientific (1 mentions) Fv1000 confocal microscope, by Olympus (1 mentions) Ab8295, by Merck Group (1 mentions) Triton x 100, by Merck Group (1 mentions)

5 protocols using ab104430

Immunohistochemical Analysis of Pediatric Brain Tumors

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Paraffin sections of brain or tumor tissues were used for hematoxylin
and eosin staining as well as immunostaining. BrdU (10 mg/kg; B5002;
Sigma-Aldrich) was injected subcutaneously in Utx^+/+and Utx^F/F Atoh1-Cre SmoM2 mice at P13. Mice were
harvested 24 hours later, and their brains were fixed in 4% PFA at 4 °C
overnight. The fixed brains were subject to subsequent double immunofluorescent
staining of BrdU and HuC/D. For systematic CD3 T cell quantification, every
5^th section throughout the tumor tissue was used to determine the
total number of T cells in each tumor sample (n=5 for
Utx^+/+ tumors and n=3 for
Utx^F/F tumors). The antibodies used were
against Ki67 (eBioscience), CD3 (ab16669, Abcam), UTX (33510, Cell Signaling
Technology), BrdU (G3G4, DHSB), HuC/D (ab184267, Abcam), microtubule-associated
protein 2 (MAP2, M9942, Sigma), NEUROD2 (ab104430, Abcam), NeuN (ABN78,
Millipore) and beta III Tubulin (TUBB3, ab78078, Abcam). Bright-field images
were acquired using a Hamamatsu Nanozoomer 2.0 HT whole slide scanner at the
University of Texas Southwestern Medical Center Whole Brain Microscopy Facility.
Fluorescent images were acquired by a confocal system (Zeiss LSM710), and three
images were taken per tumor at 200x magnification and processed using
ImageJ.

Yi J., Shi X., Xuan Z, & Wu J. (2020). Histone Demethylase UTX/KDM6A Enhances Tumor Immune Cell Recruitment, Promotes Differentiation and Suppresses Medulloblastoma. Cancer letters, 499, 188-200.

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Immunolabeling of Cerebral Organoid Slices

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Cerebral organoid slices were fixed in 4% PFA for 20 minutes. Slices were blocked and permeabilized in 5% normal donkey serum 0.25% Triton X-100 for 1 hour at room temperature. Primary and secondary antibody mix was prepared in 5% normal donkey serum 0.1% Triton X-100 and incubated overnight at 4 °C. Samples were mounted in VECTASHIELD HardSet (Vector Laboratories) and mosaics (3 × 3 tiles) were acquired with CFI Apo LWD Lambda S ×40 objective (NA 1.15; WD 0.61–0.59, Nikon). Primary antibodies used included: sheep anti-EOMES (1:200, R&D Systems AF6166), chicken anti-GFP (1:500, Abcam Ab13970), mouse anti-Sox2 (1:500, Abcam Ab79351) and rabbit anti-NeuroD2 (1:500, Abcam Ab104430).

Lindenhofer D., Haendeler S., Esk C., Littleboy J.B., Brunet Avalos C., Naas J., Pflug F.G., van de Ven E.G., Reumann D., Baffet A.D., von Haeseler A, & Knoblich J.A. (2024). Cerebral organoids display dynamic clonal growth and tunable tissue replenishment. Nature Cell Biology, 26(5), 710-718.

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NEUROD2 ChIP-Seq Reveals Differential Binding

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Previously published NEUROD2 ChIP-Seq data were used to identify differential NEUROD2 binding to genomic regions at timepoints E14.5 and P0^{26 (link),27 (link)}. These data were generated using three separate NEUROD2 antibodies (Abcam ab168932, ab104430 and ab109406). Histone ChIP-Seq datasets from E14.5 mouse embryonic cortex were produced by Bing Ren’s Laboratory, UCSD, USA^{51 (link)} (www.encodeproject.org). ChIP-Seq peaks were visualized using Trackster^{52 (link)} embedded within the Galaxy Project^{53 (link)}. Details of Closest Gene score calculations were previously described^{27 (link)}.

Guzelsoy G., Akkaya C., Atak D., Dunn C.D., Kabakcioglu A., Ozlu N, & Ince-Dunn G. (2019). Terminal neuron localization to the upper cortical plate is controlled by the transcription factor NEUROD2. Scientific Reports, 9, 19697.

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Immunofluorescent Labeling of Cryosectioned Tissue

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We fixed tissue samples in 4% paraformaldehyde for 16 h, cryoprotected them in 30% sucrose, and embedded them in optimal cutting temperature compound (Thermo Scientific). Then, 40-μm cryosections were collected on Superfrost slides (VWR) using a Leica CM3050S cryostat. Primary antibodies rabbit anti-NEUROD2 (1:500, Abcam ab104430), mouse anti-TNNT2 (1:100, Abcam ab8295), rabbit anti-GFAP (1:250, Sigma G9269) were diluted in blocking buffer containing 10% donkey serum, 0.5% Triton X-100 and 0.2% gelatin diluted in PBS at pH 7.4. Binding was revealed using an appropriate Alexa FluorTM 488, Alexa FluorTM 594, or Alexa FluorTM 647 fluorophore-conjugated secondary antibody (Life Technologies). Cell nuclei were counter-stained using DAPI (Life Technologies). Images were collected using an Olympus FV1000 confocal microscope.

Fan X., Dong J., Zhong S., Wei Y., Wu Q., Yan L., Yong J., Sun L., Wang X., Zhao Y., Wang W., Yan J., Wang X., Qiao J, & Tang F. (2018). Spatial transcriptomic survey of human embryonic cerebral cortex by single-cell RNA-seq analysis. Cell Research, 28(7), 730-745.

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Immunostaining of Neural Markers

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Mice were perfused transcardially with ice-cold 4% paraformaldehyde (in PBS). Brains were removed and post-fixed overnight at 4 °C with the same fixative. Coronal sections were cut at 50 μm thickness for all histological analyses except for dendritic reconstructions (200 μm), using a cryostat (Leica) or a sliding microtome (Microm).
Immunofluorescence experiments were performed as described before [70 (link)]. Briefly, free-floating sections were blocked and permeabilized for one hour in a blocking buffer composed of 10% Normal Goat Serum, 0.2% Triton X-100 (Sigma) in PBS. Primary antibodies, diluted in blocking solution and added overnight at 4 °C, were as follows: rabbit anti-NEUROD2 (1:500, Abcam, #ab104430), rabbit anti-TBR1 (1:1000, Abcam, #31940), rat anti-BCL11B (1:100, Abcam, #ab18465), rabbit anti-CUX1 (1:200, Santa Cruz Biotechnology, #sc13024), mouse anti-RORβ (1:200, Perseus Proteomics, #PP-N7927-00), chicken anti-GFP (1:500, Aves, #GFP-1010). For immunocytochemistry we used mouse anti-Tuj1 (1:500, Biolegend, BLE801201). Corresponding fluorescently labeled secondary antibodies (AlexaFluor, Invitrogen) were added for 2 h in blocking solution at room temperature. Hoechst was added in PBS for 10 min, and sections were mounted on microscope slides that were coversliped using Mowiol solution (Sigma).

Runge K., Mathieu R., Bugeon S., Lafi S., Beurrier C., Sahu S., Schaller F., Loubat A., Herault L., Gaillard S., Pallesi-Pocachard E., Montheil A., Bosio A., Rosenfeld J.A., Hudson E., Lindstrom K., Mercimek-Andrews S., Jeffries L., van Haeringen A., Vanakker O., Van Hecke A., Amrom D., Küry S., Ratner C., Jethva R., Gamble C., Jacq B., Fasano L., Santpere G., Lorente-Galdos B., Sestan N., Gelot A., Giacuzz S., Goebbels S., Represa A., Cardoso C., Cremer H, & de Chevigny A. (2021). Disruption of NEUROD2 causes a neurodevelopmental syndrome with autistic features via cell-autonomous defects in forebrain glutamatergic neurons. Molecular Psychiatry, 26(11), 6125-6148.

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