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Ab24574

Manufactured by Abcam
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Ab24574 is a lab equipment product offered by Abcam. It is a device used for a specific laboratory function. Details on the core function of this product are not available at this time.

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Lab products found in correlation

Mab374, by Merck Group (1 mentions) F3165, by Merck Group (1 mentions) Ab7671, by Abcam (1 mentions) Ab167453, by Abcam (1 mentions) Prolong gold mounting media, by Thermo Fisher Scientific (1 mentions) Sc 11415, by Santa Cruz Biotechnology (1 mentions) Gfp 1020, by Aves Labs (1 mentions) Alexa fluor conjugates, by Thermo Fisher Scientific (1 mentions) Mouse anti β actin ab8226, by Abcam (1 mentions)

5 protocols using ab24574

1

Antibody Characterization for Immunoblotting and Immunocytochemistry

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The following antibodies were used for immunoblotting and immunocytochemistry: FLAG (mouse, Sigma F3165, 1:500; RRID:AB_259529), GFP (rabbit; Synaptic Systems 132 002, 1:1000; RRID:AB_887725), GAPDH (mouse; Millipore MAB374, 1:5000; RRID:AB_2107445), GM130 (mouse; BD Biosciences 610822, 1:1000; RRID:AB_398141), mCherry (rabbit; Abcam ab167453, 1:500; RRID:AB_2571870), Mint2 (rabbit; Sigma M3319, 1:1000; RRID:AB_477178), alpha 1 Sodium Potassium ATPase antibody (rabbit; Abcam ab7671, 1:200; RRID:AB_306023), Neurexin-1 (rabbit; Synaptic Systems 175 103, 1:1000; RRID:AB_10697816), SMI-312 (mouse; Abcam ab24574, 1:1000; RRID:AB_448151), Synapsin (rabbit; kind gift from Dr. Thomas S üdhof, P610, 1:500), Synaptobrevin (mouse; Synaptic Systems 104 211, 1:1000; RRID:AB_2619758), alpha-tubulin (mouse; Cell Signaling 3873, 1:1000; RRID:AB_1904178).
Lin A.Y., Henry S., Reissner C., Neupert C., Kenny C., Missler M., Beffert U, & Ho A. (2019). A rare autism-associated MINT2/APBA2 mutation disrupts neurexin trafficking and synaptic function. Scientific Reports, 9, 6024.
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2

Neuromuscular Junction Quantification

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The triceps surae muscle group was isolated and dissected from the right leg of a PFA-perfused mouse, immersed in 20% sucrose-PBS mixed with OCT (1:2), pinned at its physiological resting length, flash frozen in dry-ice-cooled isopentane, and mid-point of the muscle was sectioned transversely with a cryostat on to slides at 20 μm thickness. The tissue was air dried overnight, blocked for 1 hour at room temperature in PBS containing 0.3% Triton X-100 and 2.5% bovine serum albumin (BSA), incubated overnight at 4°C in rabbit anti-neurofilament SMI-312 (1:1000; ab24574, Abcam, San Francisco, CA) diluted in blocking solution, washed several times with PBS, incubated overnight at 4°C in goat anti-rabbit IgG secondary antibody conjugated to Alexa Fluoro 488 (1:500; ThermoFisher) and rhodamine conjugated α-bungarotoxin (1:100; ThermoFisher) diluted in blocking solution, washed several times in PBS, and then mounted in ProLong Gold mounting media (ThermoFisher). Images were acquired with Leica DM2500 LED upright microscope as described above for MNs. The percentage of innervated neuromuscular junctions was determined based on the co-localization of the pre-synaptic (neurofilament) and postsynaptic (α-bungarotoxin) markers.
Franz C.K., Puritz A., Jordan L.A., Chow J., Alberto Ortega J., Kiskinis E, & Heckman C.J. (2018). Botulinum Toxin Conditioning Enhances Motor Axon Regeneration in Mouse and Human Preclinical Models. Neurorehabilitation and neural repair, 32(8), 735-745.
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3

Immunofluorescent Labeling of Neural Markers

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NG2 – rabbit polyclonal IgG anti-CSPG4 (55027-1-AP, lot#09000034, Protein Tech Group Inc., Rosemont, IL, United States) raised against a synthetic peptide corresponding to human CSPG4/NG2, GenBank accession # NM_001897.
Brevican – rabbit polyclonal IgG anti-brevican (ab106615, lot#GR163248-6, Abcam, Cambridge, MA, United States), affinity purified, raised against a synthetic peptide corresponding to amino acids 539–588 (PTETLPTPRE RNLASPSPST LVEAREVGEA TGGPELSGVP RGESEETGSS) of Human Brevican (NP_940819).
SMI-312 – mouse monoclonal IgG1 anti-SMI312 (ab24574, lot# B226113), Abcam, Cambridge, MA, United States) recognizing pan-neuronal neurofilaments commonly used as a marker for axons (Ulfig et al., 1998 (link)).
Pantazopoulos H., Hossain N.M., Chelini G., Durning P., Barbas H., Zikopoulos B, & Berretta S. (2022). Chondroitin Sulphate Proteoglycan Axonal Coats in the Human Mediodorsal Thalamic Nucleus. Frontiers in Integrative Neuroscience, 16, 934764.
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4

Immunofluorescence Staining of Cultured Cortical Neurons

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Cortical neurons were isolated and cultured as described above. Upon fixation with 4% paraformaldehyde, cells were permeabilized using 1% Triton X‐100 and blocked in 3% BSA in PBS. Antibody incubation was performed overnight at 4°C. The following antibodies were used for immunofluorescence: chicken anti‐GFP (GFP‐1020, Aves Labs Inc., 1:1,000), mouse anti‐SMI‐312 (ab24574, Abcam, 1:1,000), rabbit anti‐TOMM20 (sc‐11415, Santa Cruz Biotechnology Inc., 1:1,000), guinea pig anti‐vGat (131004, Synaptic Systems, 1:1,000), mouse anti‐vGlut1 (135311, Synaptic Systems, 1:1,000). Secondary antibodies were purchased from Jackson ImmunoResearch.
Sprenger H., Wani G., Hesseling A., König T., Patron M., MacVicar T., Ahola S., Wai T., Barth E., Rugarli E.I., Bergami M, & Langer T. (2018). Loss of the mitochondrial i‐AAA protease YME1L leads to ocular dysfunction and spinal axonopathy. EMBO Molecular Medicine, 11(1), e9288.
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5

Characterization of EFA6A Antibody

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Guinea pig polyclonal EFA6A (1626; 1:400) antibody was a gift from Prof. Eunjoon Kim (Daejeon, South Korea) and was previously characterised (Choi et al., 2006 (link)) and used in independent studies (Raemaekers et al., 2012 (link); Sannerud et al., 2011 (link)). Other primary antibodies were: mouse anti-neurofascin clone A12/18, NeuroMab (RRID:AB_10671311), 1:200; rabbit anti-Rab11 71-5300, Thermo, 1:50; rabbit anti-ARF6 ab77581, Abcam, 1:100; rabbit anti-GST ab19256, Abcam, 1:400; mouse anti-FLAG ab18230, Abcam, 1:750; mouse anti-β-actin ab8226, Abcam, 1:1000; rabbit anti-tRFP ab233, Evrogen, 1:1000; anti-integrin ß1 clone EP1041Y, 04-1109, Millipore, 1:100; and anti-pan-axonal neurofilaments mouse monoclocal SMI312, ab24574, Abcam, 1:800. ACAP1 was detected with goat anti-centaurin β1 (ab15903, Abcam) at 1:50. Secondary antibodies were Alexa Fluor conjugates from Thermo used at 1:1000. Secondary antibodies for western blotting were horseradish peroxidase (HRP) conjugates from GE Life Sciences used at 1:10,000.
Eva R., Koseki H., Kanamarlapudi V, & Fawcett J.W. (2017). EFA6 regulates selective polarised transport and axon regeneration from the axon initial segment. Journal of Cell Science, 130(21), 3663-3675.
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