Arl13b

Manufactured by Proteintech

Sourced in United States

ARL13B is a protein that is a member of the Arf-like (Arl) small GTPase family. It functions as a regulator of primary cilium formation and maintenance.

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Lab products found in correlation

Ift88, by Proteintech (8 mentions) Acetylated α tubulin, by Merck Group (6 mentions) γ tubulin, by Merck Group (4 mentions) Ift81, by Proteintech (3 mentions) Acetylated tubulin, by Merck Group (3 mentions) Ift140, by Proteintech (3 mentions) Alexa fluor 488, by Thermo Fisher Scientific (3 mentions) Gapdh, by Novus Biologicals (3 mentions) Anti acetylated α tubulin, by Merck Group (2 mentions) Pericentrin, by Abcam (2 mentions) Confocal 810, by Zeiss (2 mentions) Flag tag (flag), by Merck Group (2 mentions) Smoothened, by Santa Cruz Biotechnology (2 mentions) Gt335, by Adipogen (2 mentions) Phospho jnk1 2, by Thermo Fisher Scientific (2 mentions) β actin, by Cell Signaling Technology (2 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (2 mentions) β actin, by Merck Group (2 mentions) Alexa fluor 568, by Thermo Fisher Scientific (2 mentions) Lsm 510, by Zeiss (2 mentions)

Close spelling variants of this product

Anti-Arl13b Anti-ARL13B (17711-1-AP) Rabbit anti-ARL13B antibody Anti-Arl13b antibody Rabbit anti-ARL13b (17711-1-AP; Arl13b (17711-1-AP) Rabbit anti-ARL13B ARL13B antibody Arl13B rabbit polyclonal antibody (17711-1-AP)

41 protocols using arl13b

Immunofluorescence Antibody Characterization

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Antibodies used were against the following proteins: acetylated tubulin (Sigma-Aldrich, Cat# T6793, RRID:AB_477585, used 1:1000 for immunofluorescence, methanol fixation); ARL13B (Proteintech, Cat# 17711–1-AP, RRID:AB_2060867, 1:1000 for immunofluorescence, methanol fixation); ARL13B (Proteintech, Cat# 66739-1-Ig, RRID:AB_2882088, 1:1000 for immunofluorescence, methanol or 4% PFA fixation); GAPDH (Proteintech, Cat# 60004-1-Ig, RRID:AB_2107436, 1:5000 for immunoblotting); GFP (BioLegend, Cat# 902601, RRID:AB_2565021, 1:5000 for immunoblotting, methanol fixation); IFT88 (Proteintech, Cat# 13967-1-AP, RRID:AB_2121979, 1:300 for immunofluorescence, 4% PFA fixation); IFT140 (Proteintech, 17460-1-AP, RRID:AB_2295648, 1:100 for immunofluorescence); RPGRIP1L (Proteintech, Cat# 55160–1-AP, RRID:AB_10860269, 1:200 for immunofluorescence, Methanol fixation); and SMO (Santa Cruz Biotechnology, Cat# sc-166685, RRID:AB_2239686, 1:100 for immunofluorescence, 4% PFA fixation).

Shak C., Vuolo L., Uddin B., Katoh Y., Brown T., Mukhopadhyay A.G., Heesom K., Roberts A.J., Stevenson N., Nakayama K, & Stephens D.J. (2022). Disease-associated mutations in WDR34 lead to diverse impacts on the assembly and function of dynein-2. Journal of Cell Science, 136(5), jcs260073.

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Immunohistochemical Analysis of ACIII and ARL13B

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Parallel free-floating sections were subjected to endogenous peroxidase blocking with 1% H₂O₂ in PBS, followed by treatment with blocking buffer (1% fetal bovine serum (FBS) in PBS and 0.3% Triton X-100 for 30 min) and incubation with primary anti-acetylcholine III (ACIII, 1:500, #sc-588, Santa Cruz, CA) overnight. Immunohistochemical staining of the tissue sections were performed using the avidin–biotin peroxidase complex (ABC) method described previously [11 (link), 18 (link)]. HT22 hippocampal cells were immunoreacted for ADP ribosylation factor like GTPase 13B (ARL13B, 1:1000, #17711-1-AP, Proteintech). All immunoreactions were incubated with Cy3-conjugated anti-rabbit secondary antibody and counterstained with DAPI.

Baek H., Shin H.J., Kim J.J., Shin N., Kim S., Yi M.H., Zhang E., Hong J., Kang J.W., Kim Y., Kim C.S, & Kim D.W. (2017). Primary cilia modulate TLR4-mediated inflammatory responses in hippocampal neurons. Journal of Neuroinflammation, 14, 189.

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Immunofluorescence Analysis of Primary Cilia

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Cells were seeded onto Poly-L-Lysine coated coverslips in 24-well plates. At desired experimental time points, cells were fixed with 4% paraformaldehyde in phosphate-buffered saline containing 0.1% Triton x-100 for 10 minutes at room temperature. Cells were blocked in 2% Bovine Serum Albumin (Sigma) for 1 hour at room temperature. Cells were then incubated with primary antibody diluted in blocking solution overnight at 4°C. Primary antibodies used include: acetylated alpha-tubulin (Sigma), ARL13B (Proteintech), IFT-81 (Proteintech), γ-tubulin (Sigma), PPARγ (Cell Signaling), and CEBPα (Cell Signaling). Cells were washed and incubated with secondary antibody, AF488 or AF542 (Life Technologies) for 1 hour at room temperature. To visualize lipid droplets, slides were incubated with BODIPY (Fisher) for one hour at room temperature. Coverslips were inverted and mounted onto slides using DAPI Fluoromount-G (Electron Microscopy Services, 17984–24). Cells were visualized and imaged using a Nikon80i microscope attached to a Nikon DS-Fi1 camera.

Jacobs D.T., Allard B.A., Pottorf T.S., Silva L.M., Wang W., Al-Naamani A., Agborbesong E., Wang T., Carr D.A, & Tran P.V. (2019). Intraflagellar-transport A dysfunction causes hyperphagia-induced systemic insulin resistance in a pre-obese state. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 34(1), 148-160.

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Embryonic Mouse Brain Histology Protocol

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Embryonic mouse brains were dissected in PBS and fixed overnight in 4% paraformaldehyde (PFA) at 4°C. Postnatal animals were perfused with 0.9% saline and PFA, and their brains were dissected and fixed in PFA overnight at 4°C. Histology, immunostaining, and TUNEL assay were performed as described previously (Lavado et al., 2018 (link)). Luxol blue and cresyl violet staining was performed using the Kluver-Barrera method (EMS #26681). The primary antibodies used were: YAP/TAZ (Cell Signaling #8418, 1:500), YAP (Cell Signaling #14074, 1:250), TAZ (Cell Signaling #83669, 1:100 with the Tyramide SuperBoost Kit, Invitrogen #B40922), SOX2 (Santa Cruz Biotechnology #SC17320, 1:100), SOX2–Alexa-647 (BD Biosciences #562139, 1:100), TBR2 (Thermo #14-4875-82, 1:250), TBR2 (Abcam ab23345, 1:250), TBR1 (Abcam ab31940, 1:1000), CUX1 (Santa Cruz #SC13024, 1:250), CTIP2 (Abcam ab18465, 1:1000), PAX6 (BioLegend #901301, 1:500), BrdU/IdU (BD Biosciences #347580, 1:50), BrdU (Abcam ab152095, 1:500), Ki67 (Vector laboratories #VP-RM04, 1:500), ZO1–Alexa-488 (Thermo #339188, 1:100), Nestin (R&D systems #AF2736, 1:100), ARL13b (ProteinTech #17711-1-AP, 1:250), β-Catenin–Alexa-647 (Cell Signaling #4627, 1:100), and S100β (Sigma #52532, 1:100).

Lavado A., Gangwar R., Paré J., Wan S., Fan Y, & Cao X. (2021). YAP/TAZ Maintain the Proliferative Capacity and Structural Organization of Radial Glial Cells during Brain Development. Developmental biology, 480, 39-49.

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Immunocytochemistry of Medaka Embryonic Eyes

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Eyes from medaka embryos were processed for immunocytochemistry as described (Conte et al, 2010b (link)). Embryos were subjected to anesthesia before fixation at stage 40 by 2 h of incubation in 4% paraformaldehyde, 2× phosphate‐buffered saline (PBS), and 0.1% Tween‐20 at room temperature (RT). Samples were rinsed three times with PTW 1× (PBS 1×, 0.1% Tween‐20, pH 7.3) and then incubated overnight (ON) in 15% sucrose/PTW 1× at 4°C, and then again incubated overnight in 30% sucrose/PTW 1× at 4°C. Cryosections of the larvae were processed for immunostaining. Sections were rehydrated in PTW 1×. Blocking solution containing 10% FBS/PTW 1× was applied for 1 h at RT. Primary antibodies (GFP Invitrogen A‐6455, 1:500, Rhodopsin Thermo Fisher MA5‐11741, 1:100) were diluted in 5% FBS/PTW 1×, and sections were incubated ON at 4°C. ARL13B (1:500; #17711‐I‐AP, Proteintech). Thus, samples were washed three times with PTW 1× and incubated with secondary antibody Alexa 488 anti‐rabbit IgG (1:1,000, Invitrogen A‐11037) and Alexa 594 anti‐mouse IgG (1:1,000, Invitrogen A‐11032). Nuclei were stained with DAPI 1:500 in PBS for 10 min at RT. Finally, sections were washed three times with PTW 1× and mounted with PBS/glycerol solution.

Chiuso F., delle Donne R., Giamundo G., Rinaldi L., Borzacchiello D., Moraca F., Intartaglia D., Iannucci R., Senatore E., Lignitto L., Garbi C., Conflitti P., Catalanotti B., Conte I, & Feliciello A. (2023). Ubiquitylation of BBSome is required for ciliary assembly and signaling. EMBO Reports, 24(4), e55571.

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Immunofluorescence and Immunoblotting Antibodies

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Antibodies were from the following sources: N-KDM3A (12835; Proteintech), C-KDM3A (NB100-77282; Novus Biologicals), anti–acetylated α-tubulin (T7451; Sigma-Aldrich), γ-tubulin (GTU-88; Sigma-Aldrich), IFT88 (13967-1-AP; Proteintech), IFT81 (11744-1-AP; Proteintech), GAPDH (5019A-2; ImGENEX), ARL13B (17711-1-AP; Proteintech), DDK tag (TA50011; OriGene), and Halo (G9281; Promega). Secondary antibodies for immunofluorescence and immunoblotting are Fab′2 IgG Alexa Fluor from Molecular Probes and HRP conjugated from EMD Millipore and Sigma-Aldrich.

Yeyati P.L., Schiller R., Mali G., Kasioulis I., Kawamura A., Adams I.R., Playfoot C., Gilbert N., van Heyningen V., Wills J., von Kriegsheim A., Finch A., Sakai J., Schofield C.J., Jackson I.J, & Mill P. (2017). KDM3A coordinates actin dynamics with intraflagellar transport to regulate cilia stability. The Journal of Cell Biology, 216(4), 999-1013.

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Immunostaining and Immunoblotting Antibody Panel

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The primary antibodies used for immunostaining and immunoblotting were as follows: AC3 (Santa Cruz, #sc-588), ARL13B (Proteintech, #17711-1-AP), BrdU (Novus, #NB500-169), β-END (Phoenix Pharmaceuticals, #H-022-33 or Abcam, #Ab32893), IFT88 (Proteintech, #13967-1-AP), LepRb (R&D Systems, #AF497), LAMP1 (BD Pharmingen, #553792), LC3B (Abcam, #Ab51520), MAP2 (Abcam, #Ab5392), Neurofilament (Biolegend, #837904), NPY (Abcam, #Ab6173), p62 (Abcam, #Ab91526), and TFEB (Bethyl Laboratories, #A303-673A).

Lee C.H., Song D.K., Park C.B., Choi J., Kang G.M., Shin S.H., Kwon I., Park S., Kim S., Kim J.Y., Dugu H., Park J.W., Choi J.H., Min S.H., Sohn J.W, & Kim M.S. (2020). Primary cilia mediate early life programming of adiposity through lysosomal regulation in the developing mouse hypothalamus. Nature Communications, 11, 5772.

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Immunofluorescence Analysis of Primary Cilia

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Cells were cultured in 4-well chamber slides (LabTek). After serum starvation, cells were washed twice with PBS and fixed in PFA for 10 min and then permeabilized with 0.1% Triton-X-100 in PBS for 15 min at room temperature. Cells were then blocked in 10% goat serum for 1 h at room temperature. Primary antibodies were diluted in PBS containing 1% serum and incubated overnight at 4 °C. Antibodies used were as follows: ARL13B (Proteintech, 1:100); acetylated α-tubulin (Sigma T6793, 1:2000); pericentrin (Abcam, 1:2000). DAPI was used to stain the nuclei. Detection was performed with secondary Alexa Fluor 488/568 antibodies (Invitrogen, 1:1000). Images were captured on Zeiss Confocal 810. Images were collected with 1024 × 1024 pixel definition and Z-sections were taken at 0.5-µm step size. Max projections of the Z-stacks were used for primary cilium counting in ImageJ (NIH).

Duran I., Taylor S.P., Zhang W., Martin J., Qureshi F., Jacques S.M., Wallerstein R., Lachman R.S., Nickerson D.A., Bamshad M., Cohn D.H, & Krakow D. (2017). Mutations in IFT-A satellite core component genes IFT43 and IFT121 produce short rib polydactyly syndrome with distinctive campomelia. Cilia, 6, 7.

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Measuring Primary Cilia Length

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Cilia length were measured by direct immunofluorescence for cilia marker with anti-Acetylated α-tubulin or Arl13b staining. The cells were fixed for 10 minutes (4% paraformaldehyde/2% sucrose in PBS) and permeabilized for 5 minutes (10% triton X-100). Acetylated α-tubulin (1:10,000 dilution, Sigma Aldrich, St. Louis, MO) or Arl13b (1:50 dilution, Proteintech, Rosemont, IL) and fluorescein isothiocyanate (FITC)-conjugated (1:1000 dilution, Vector Labs Burlingame, CA) antibodies were each incubated with the cells for 1 hour at 37°C. Microscope slides were then mounted with DAPI (Southern Biotech, Birmingham, AL) hard set mounting media. Nikon Eclipse Ti-E inverted microscope with NIS-Elements imaging software (version 4.30) was used to capture images of primary cilia. Automated image acquisition was conducted in 100X magnification fields. Cilia length analysis was followed a standard calculation as previously described^{56 (link)}.

Nam Y.W., Pala R., El-Sayed N.S., Larin-Henriquez D., Amirrad F., Yang G., Rahman M.A., Orfali R., Downey M., Parang K., Nauli S.M, & Zhang M. (2022). Subtype-Selective Positive Modulation of KCa2.3 Channels Increases Cilia Length. ACS Chemical Biology, 17(8), 2344-2354.

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Immunofluorescence and Western Blot Antibodies

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Antibodies used in this study consist of those targeting BBS3 (Santa Cruz Biotechnology, Cat# sc-390021), BBS2 (Santa Cruz Biotechnology, Cat# sc-365355 and Proteintech, Cat# 11188-2-AP), acetylated α-tubulin (Santa Cruz Biotechnology, Cat# sc-23950), ADP ribosylation factor like GTPase 13B (ARL13B, Proteintech, Cat# 17711-1-AP), β-actin (Proteintech, Cat# 66009-1-Ig), NPY₂R (Neuromics, Cat# RA14112), POMC (Phoenix Pharmaceuticals, INC, Cat# H-029-30), DRP1 (BD Transduction Laboratories, Cat# 611113), and phospho-DRP1 (Ser616, Cell Signaling, Cat# 3455). For Western blotting, primary antibodies were diluted 1:1000 in 5% bovine serum albumin (BSA). For immunofluorescent staining, primary antibodies were diluted 1:250.

Guo D.F., Williams P.A., Laule C., Seaby C., Zhang Q., Sheffield V.C, & Rahmouni K. (2023). POMC Neuron BBSome Regulation of Body Weight is Independent of its Ciliary Function. Function, 5(1), zqad070.

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