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We narrowed to 37 results for: snap-tag

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  1. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    Published
    April 16, 2024, 1:15 p.m.
    ...became known as SNAP-tag. There is also a fast-reacting version of SNAP-tag called SNAPf. eDHFR: The Virginia...Characterization of the Self-Labeling Protein Tags HaloTag7, SNAP-tag, and CLIP-tag. Biochemistry, 60(33), 2560–2575...than short peptide tags like tetracysteine (Jing & Cornish, 2011). HaloTag and SNAP-tag are the most popular...application, SNAP-tag, CLIP-tag (both 182 aa), and eDHFR (159 aa) are smaller than HaloTag (295 aa) – for...on resorufin). SNAP-tag: In 2003, Kai Johnsson’s lab developed a self-labeling fusion tag based on the ...Although based on the same original enzyme, SNAP-tag and CLIP-tag show good specificity for their own ligands...genetic tag on the protein of interest. Table 1: Summary of popular self-labeling tag systems Tag Size...

  2. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    Published
    June 29, 2017, 1:06 p.m.
    ...resistance genes, selectable markers, promoters, origins, tags, and certain ORFs. Their sophisticated detection...powered by GSL Biotech's SnapGene Server Software. With the backing of SnapGene’s sequence viewer software... Overview of SnapGene powered plasmid maps at Addgene  Three factors make our SnapGene powered plasmid...the features you need quickly. SnapGene’s Feature Library - SnapGene’s software seamlessly identifies ...example below. Our old map is on the left while the SnapGene powered map is on the right (click here to see...t detected by our in-house software. In fact, SnapGene’s feature library includes nearly 3x as many features...identify. Improved Restriction Enzyme Handling - SnapGene’s software positions restriction enzyme annotations...

  3. Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process

    Type
    Blog Post
    Published
    Feb. 1, 2018, 3:07 p.m.
    ...markers, tags, origins of replication, and fluorescent proteins that are already included in Snapgene’s extensive...through our Snapgene powered maps and sequence analysis tools.     The main ways we use SnapGene for plasmid...version of Snapgene to visualize the sequence and look for features. The first thing that the Snapgene software...software as the desktop version of Snapgene. When you download a Snapgene file directly from our website,...website, you can open it with either the Snapgene viewer or Snapgene Desktop programs to analyze the sequence... plasmids. We’ve also talked about how our new Snapgene generated maps provide improved feature detection...with an easy to use interface. We regularly use Snapgene for our quality control process because of its...

  4. Hot Plasmids: Winter 2025

    Type
    Blog Post
    Published
    March 11, 2025, 1:15 p.m.
    ...Twelvetrees Lab took advantage of the self-labeling tags SNAPf and CLIPf. These tags rely on synthetic dye...double-tagged constructs (including "CLAP", combining an N-terminal CLIPf tag and a C-terminal SNAPf tag;...10.1101/2024.02.11.579803).  CLIP, CLAP, SNAP! Self-labeling tags for single-molecule biophysics By Mike...Figure 2: CLIP-SNAP fusion proteins for single-molecule FRET. A) Schematic of a CLIP-SNAP fusion protein...dicots Self-labeling tags for single-molecule biophysics Fast and responsive voltage indicator ONE-GO biosensors...constructs for tagging the kinesins KIF5A and KIF5B as well as backbones for generating other SNAPf-, CLIPf-...CLIPf-, and CLAP-tagged proteins for mammalian cell expression. These plasmids will be useful not only for...

  5. Identifying Sequence Elements with SnapGene's Feature Database

    Type
    Blog Post
    Published
    Dec. 21, 2017, 2:06 p.m.
    ... who uses SnapGene or the free SnapGene Viewer companion product. If you don’t use SnapGene yet, you can...guest bloggers Aline and Benjamin Glick from SnapGene. SnapGene was created to meet a need. While there were...approach is SnapGene’s algorithm for detecting common features. This algorithm enables one of SnapGene’s most... us at snapgene.com. Many thanks to our guest bloggers Aline and Benjamin Glick from SnapGene!  Ben Glick...Addgene Blog Read about our SnapGene powered plasmid maps Learn how we use SnapGene during the plasmid quality...properties of the DNA molecules they were using. SnapGene was created to alleviate these problems through...make specialized controls available when needed. SnapGene has been engineered to be easy and enjoyable to...

  6. #DeckTheLab to Win a Tasty Winter Treat This Holiday Season

    Type
    Blog Post
    Published
    Nov. 30, 2015, 3:30 p.m.
    ...wintertime festivities! Help us spread some cheer by snapping a photo of you and your labmates celebrating the... the holidays and tweeting it with the tags #DeckTheLab and @Addgene. Christmas, Hanukkah, ugly sweaters...you name it, we want to see it.      All photos tagged with #DeckTheLab throughout the month of December...

  7. 'Tis the Season to #DeckTheLab

    Type
    Blog Post
    Published
    Dec. 2, 2016, 3:30 p.m.
    ... Why #DecktheLab? Help us spread some cheer by snapping a photo of you and your labmates celebrating the... the holidays and tweeting it with the tags #DeckTheLab and @Addgene. We’d love to see how you’re decorating...Festivus— you name it, we want to see it! All photos tagged with #DeckTheLab throughout the month of December...

  8. HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides

    Type
    Blog Post
    Published
    Oct. 10, 2019, 12:41 p.m.
    ...other fluorescent proteins and tags including mCherry, HaloTag and SNAP-tag for multi-color imaging. Find...observed colocalization of GFP tagged HA frankenbody with an mCherry, 4xHA tagged histone protein (H2B) in ...could also bind a 1xHA tagged histone protein as well as a mitochondrial protein tagged with spaghetti monster... the 10xHA tag increased the fluorescent signal to noise ratio five fold over the 1xHA tag.  Lastly, the...use GFP to image nascent proteins. In addition GFP tags are large, dim, and permanently attached to a protein...limitations, scientists have co-opted antibodies for tagging purposes. Different antibodies bind different protein... called epitopes and scientists can use these to tag their proteins of interest. Antibody based probes...

  9. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    Published
    Jan. 24, 2019, 2:24 p.m.
    ... with containing chemical tags such as the SNAP-tag, Halo-tag, and CLIP-tag (Kohl et al., 2014) (Addgene... et al. "Ultrafast tissue staining with chemical tags." Proceedings of the National Academy of Sciences111.36... circuits with fluorescent proteins and chemical tags." eLife 7 (2018): e40350. PubMed PMID: 30454553....

  10. Hot Plasmids - February 2022

    Type
    Blog Post
    Published
    Feb. 22, 2022, 2:15 p.m.
    ...frankebodies includes mEGFP, mRuby2, iRFP713, SNAP-tag, and HaloTag fusions, while the anti-HA frankenbodies... small epitope tag (FLAG is eight amino acids, HA is nine), multiple repeats of the tag can recruit multiple...b) Schematic of anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of...blotting, and more. Given the popularity of FLAG and HA tags, you may not even need to clone new constructs to.... The proteins, which are purified via histidine tags, are optimized for SDS-PAGE gels and can be detected...frankenbodies include mEGFP, mCherry, and HaloTag. Figure 1: (a) Anti-FLAG frankenbody (FB) construction from...editing within a single expression cassette is advantageous to Cas9’s larger expression requirements (Figure...

  11. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    Published
    Oct. 12, 2017, 2:26 p.m.
    ...T2A-TurboGFP-PEST), and small epitope tags (HA, Myc, Strep tag II, AviTag, HaloTag, SpyTag). The Foerstemann lab developed...types of fluorescent (EGFP, tagRFP, mCherry) and nonfluorescent (Halo, SNAP) tags, a selectable marker (floxed... scarless tag insertion.   In addition to the obvious advantages SapTrap offers for tagging a single genetic...between the tag and homology arms). Digestion of the donor plasmids with SapI allows the tag, selectable... inserting a tag at multiple target sites. SapTrap vectors can be easily modified to tag hundreds of genes...insertion tagging), developed by the Hornung Lab, allows users to easily create C-terminal tag fusions ...endogenous protein tagging, please visit our website CRISPR/Cas Plasmids - Protein Tagging. Beyond simply...

  12. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    Published
    Sept. 8, 2016, 2:30 p.m.
    ...concept include: Self-labeling tags (e.g., SNAP-tag, HaloTag, TMP-tag) – These widely used systems consist...strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion with your favorite protein...fluorophore-contianing “ligand” that binds to the tag. Like most good ideas in biological imaging, the ...genetically encoded tetracysteine (Cys4) peptide tag could be used to label proteins in cells (Figure ...consist of a genetically encoded enzyme variant tag that reacts specifically and irreversibly with a small... attachment of a fluorophore ligand to a peptide tag (Figure 1c). Click chemistry (e.g., transcyclooctene–tetrazine...paclitaxel. Unlike other systems, no genetically encoded tag is required as the small-molecule binding motif targets...

  13. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...C-terminal fusion of a tetracysteine tag. In addition, since these tags are so small, one can insert them... lure the ribosome into inserting a fluorophore-tagged amino acid at a stop codon (often amber i.e. UAG...termination at stop codons, expression levels of these tagged proteins are often very low. In addition, a major...which can be carried out using Addgene plasmids. His-tag labeling reagents You may already have a FRET acceptor-binding...: you just don’t know it yet! The poly-histidine tag often used for protein purification is a reliable...for protein purification: the NTA binds to the His tag on your protein and the coupled Cy fluorophore acts...well as by extending the length of the histidine tag from 6 to 10 residues. While these Cy/NTA conjugates...

  14. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    Published
    March 21, 2024, 1:15 p.m.
    ... would be SNAP-tag® and HaloTag®. These consist of a genetically encoded enzyme variant tag that covalently...several advantages to using these self-labeling proteins over traditional fluorescent protein tags, such...endogenous target, and thus no genetically encoded tag is required. These dyes can also be attached to an...ligands have more flexibility due to the affinity tag they typically bind to. Stability of fluorescence...experimental labeling. Size is also a factor. Bulkier tags, such as GFP, may interfere with protein expression...fluorescent experiments, such as using fluorescently tagged ligands or intermolecular FRET for ligand-receptor...should possess high specificity to ensure accurate tagging. In Fluorescence Resonance Energy Transfer (FRET...

  15. Important Considerations When Using AAVs

    Type
    Blog Post
    Published
    June 13, 2017, 2:30 p.m.
    ... storage, make > 25 μl aliquots in 0.5 ml tubes, snap freeze in liquid nitrogen or a dry ice/ethanol bath...them. There are suspected cases of insertional mutagenesis in humans (1) and AAVs are non-enveloped viruses... Russell, David W. "AAV vectors, insertional mutagenesis, and cancer." Molecular Therapy 15.10 (2007):...

  16. Transferable Skills Guide: Teamwork

    Type
    Blog Post
    Published
    Nov. 28, 2017, 2 p.m.
    ...identify as one.  But some time around 10th grade, I snapped out of the phase of my life that had me listening...the first place? Let’s think about some of the advantages of team-based work: Nobody knows everything anymore...

  17. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    Published
    Oct. 27, 2015, 3 p.m.
    ...and gram negative bacteria periplasm. Efforts to tag endogenous genes for proteins in these compartments...Munro, Sean, and H. R. Pelham. "Use of peptide tagging to detect proteins expressed from cloned genes:...Avaliable at Addgene Read Our Blog Post on Protein Tags Read Other Posts on Fluorescent Proteins Browse ...our guest bloggers Erik L. Snapp and Lindsey M. Costantini! Erik Lee Snapp received his PhD in Molecular... post was contributed by guest bloggers Erik L. Snapp and Lindsey M. Costantini. "You underestimate the...FP-fusion proteins functionally relative to the untagged protein of interest. To avoid the accumulation...research was conducted in the laboratory of Erik Lee Snapp and focused on optimizing fluorescent proteins for...

  18. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    Published
    July 2, 2020, 12:40 p.m.
    ... Replace the biotinalatable tag on dCas9 with a BioTAP-tag. The BioTAP-tag is a 69 aa long biotinylation... lentiviruses delivering 1) a BioTAP-tagged dCas9 and an eGFP tag, and 2) gRNAs Number of loci targeted...normally expressed in eukaryotic cells. Using this tag reduces the number of CAPTURE components that need...from three to two. Lentiviral delivery of BioTAP-tagged dCas9 and gRNAs. Using lentiviral delivery of the...system Stable cell line expressing 1) FLAG-biotin-tagged dCas9, 2) BirA, and 3) one or more gRNAs Two separate...et al., 2018). Think of 3C methods as taking a snapshot of chromatin looping with the photo “developed...

  19. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    Published
    April 19, 2016, 2:30 p.m.
    ...solution or something in between, you are using a tag with a serious drawback. The standard EGFP plasmid... of a POI. As an investigator, you want a fusion tag to be inert, to not produce artifacts in your experiments...Many thanks to our guest blogger Erik L. Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by guest blogger Erik L. Snapp. Stop using EGFP/GFP for fusion proteins! Despite...oligomers such as DsRed or high affinity FPs such as TagRFP. Problem 3 can be solved, in most cases, by using...distance runner, gardner, and cook.   References 1. Snapp, E.L., et al., Formation of stacked ER cisternae... Resources on the Addgene Blog Read Professor Snapp's Post on mOX FPs Find the Best Fluorescent Protein...

  20. A Look at Addgene's QC Process

    Type
    Blog Post
    Published
    May 6, 2025, 1:15 p.m.
    ...plasmid they ordered contains the correct inserts, tags, promoters, and other critical functional components...on the information provided and vector type  Any tags, selectable markers, and fusion proteins listed ...deposited sequence(s) in SnapGene. SnapGene annotates the plasmid maps based on SnapGene’s “Common Feature” library...

  21. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    Published
    March 1, 2016, 3:30 p.m.
    ...into a vector with a large tag at the N-terminus, but you don’t have the tag already included in the vector...you can insert both the gene of interest and the tag sequences into the vector in one step without scars...In this blog post, I will go over some advantages, disadvantages, and examples of how scientists are using... common, and for good reason. They offer many advantages over the traditional restriction enzyme cloning...thereof) nor a serious time commitment. Another advantage is that this process makes it easy to generate...primer melting temperature greater than 48℃. Both Snapgene and NEB have tools that help you design primers...video gives a helpful demonstration of how to use Snapgene’s program to design primers for Gibson Assembly...

  22. Hot Plasmids: Spring 2025

    Type
    Blog Post
    Published
    June 3, 2025, 1:15 p.m.
    ... cells. Flexible tagging: Choose untagged or add a versatile N- or C-terminal HA tag. Selectable and stable...breeze. Modular by design: Key elements (promoter, tag, polyA site) are flanked by unique restriction sites...plasmid set, our bacterial plasmid set for expressing tagged proteins, and stay tuned for mammalian expression...streptavidin immunoprecipitation (IP), and tandem mass tag (TMT) mass spectrometry (MS) in HCT116 cells (colorectal...with tools like mass spectrometry, providing a snapshot of interacting proteins at a certain time and ...and scale bar are indicated. B) Specificity, as percentage of BiPVe3-dTomato and PV-positive cells out of...

  23. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...plasmid.     Confirming tags and fusion proteins Finally, we confirm promoters, tags, fusion proteins, and...aligning to NCBI entry or using BLAST.   Confirming tags and fusion proteins. We’ll break down each of these...selectable markers by detecting common features using Snapgene. If we find information that differs from what...

  24. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    Published
    Oct. 18, 2018, 12:37 p.m.
    ...short sequences of up to 100 bp, such as epitope tags, design primers with template binding regions to...combining multiple primers, a C-terminal epitope tag can be moved to the N-terminus of a protein-coding... can be introduced in a gene, while adding a GFP-tag (Figure 3). Achieving multiple modifications requires... to the 5’ end of the reverse primer. Mutagenesis - Mutagenesis is performed by designing primers astride...modification, from insertions, deletions, and point mutagenesis to complex assembly of multiple genes. Primer...a sequence manager A sequence manager, such as SnapGene, can be very helpful to confirm primer design....rapid method for recombination and site-specific mutagenesis by placing homologous ends on DNA using polymerase...

  25. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    Published
    May 29, 2014, 1:29 p.m.
    ...to an epitope tag or fusion protein. If your sequencing primer was chosen to confirm a tag or fusion protein...expected ORFs, mutations or truncations. A primary advantage of blastx is that you do not have to decide on...recommend using blastp over blastx. The primary advantage is time savings but an added benefit is that blastp...other matrices to see if another would be more advantageous for your search. Timesaving Tip #3: Note that...at NGS Plasmid Quality Control Learn about our Snapgene-powered plasmid maps. Resources on Addgene.org...

  26. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    Published
    March 12, 2019, 1:37 p.m.
    ... Many thanks to our guest blogger, Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at the Janelia...Growth of new livers in culture could resolve this shortage." What is the knowledge gap you plan to address...

  27. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    Published
    Oct. 9, 2018, 12:21 p.m.
    ... Many thanks to our guest blogger, Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at theJanelia... be needed to achieve your intended next career stage and create a timeline for achieving these goals....

  28. Our Most Popular Blog Posts from 2019

    Type
    Blog Post
    Published
    Dec. 31, 2019, 2:26 p.m.
    ...Academic Job Interview by Erik Snapp Addgene depositor and guest blogger Erik Snapp breaks down how to prepare... 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins by Cary...

  29. Visualizing Translation at the Single Molecule Level

    Type
    Blog Post
    Published
    Aug. 1, 2017, 1:15 p.m.
    ...contains 24 copies of SunTag, a synthetic scaffold that can recruit GFP fused to the SunTag-specific antibody...biochemical method developed in 2009, takes a “snapshot” of all of the mRNA bound-ribosomes in a cell....Imaging by Coat protein Knock-off) instead takes advantage of the ribosome’s movement. When the ribosome ...indicating translational repression. During later stages, single RFP-labeled osk and Oskar protein were ...scFv. Each time the reporter is translated, the SunTag sites on the nascent polypeptide recruit scFV-GFP..., as the authors of these papers did. Since the SunTag-based system tethers mRNAs to the plasma membrane...Additional Resources the Addgene Blog Learn More about SunTag Catch up on Your GFP Background Choose the Brightest...

  30. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    Published
    June 30, 2016, 2:30 p.m.
    ...newly synthesized product from a site-directed mutagenesis experiment? The answer to both questions is the...to methylation if the enzyme's recognition site (TCTAGA) is preceded by GA or followed by TC. As shown ...cleave DNA. DpnI is often used for site directed mutagenesis. During this process, incorporation of a desired...illustrates how sequence visualization software (we used Snapgene here) can help you assess whether your restriction...Troubleshoot Your Sequencing Results Use REPLACR Mutagenesis to Mutate Your Plasmids Resources on Addgene.org...

  31. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    Published
    July 13, 2017, 2:30 p.m.
    ...three larval stages where larvae molt in order to continue growing in the next larval stage. After the ...the third larval stage, Drosophila will pupate, which takes about one week from the initial deposition of...through their translucent bodies. This is the ideal stage to select for females because you can be certain...Drosophila cell lines are extremely effective. Their Snapdragon tool helps you select primers to synthesize RNA...

  32. A Deep Dive into BioBuilder

    Type
    Blog Post
    Published
    Oct. 11, 2017, 1:44 p.m.
    ...and in more than 30 countries around the world. Snapshots of the BioBuilder experience With its intentionally...recognized platform in March to exhibit projects at any stage of completion, from pitch to prototype. Last year...preparing local (Massachusetts), economically disadvantaged students for paid summer internships. A summer...

  33. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    Published
    Jan. 12, 2017, 3:30 p.m.
    ...more about the Gateway cloning method and its advantages. An introduction to Gateway technology The Gateway..., etc). This video demonstrates how to use the Snapgene program to design Gateway plasmids.    ...and verify that your construct is functional. Advantages of the Gateway cloning method Compatibility and...JY, Chin CF (2015) Gateway Cloning Technology: Advantages and Drawbacks. Clon Transgen 4:138. doi:10.4172...

  34. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ...possible consequences of FP aggregation, as Erik Snapp’s blog post shows. To avoid these issues, Subach ...knowledge of protein structure and saturation mutagenesis. mK-GO matures from green to red, and the FT ...absorption spectra). Subach et al’s directed mutagenesis also produced the three separate timers shown...

  35. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    Published
    Nov. 6, 2018, 1:12 p.m.
    ...lineage, without dedifferentiating into a pluripotent stage. Cellular reprogramming The reversal of a differentiated...reprogramming [1]. The process can be divided into two stages: Dedifferentiation - Conversion of adult somatic...differentiated cells of choice. The dedifferentiation stage involves overexpression of four reprogramming factors...differentiated somatic cell to revert back to a pluripotent stage (iPSC formation) [2, 3]. The iPSCs then proliferate...reprogrammed to iPSCs successfully [10-12]. The next stage of reprogramming consists of redifferentiation of... resources More (due to additional pluripotency stage) Less Safety Less (C-MYC, a known oncogene is ...Williams, L.A., B.N. Davis-Dusenbery, and K.C. Eggan, SnapShot: directed differentiation of pluripotent stem ...

  36. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    Published
    June 12, 2018, 11:40 a.m.
    ...grows on a daily basis, the lists below provide a snapshot of relevant resources and a good jumping off point...H3.1 H3.3 HIST1H1E HIST1H2BB HIST2H2BE RBX1 STAG2 Xist Your favorite gene! SWI/SNF Genes:       ...

  37. Deep Dive: Statistical Tests (Comparisons)

    Type
    Blog Post
    Published
    March 7, 2023, 2:15 p.m.
    ...tendency – mean, median, and mode give you a summary snapshot without telling you anything about outliers or...dependent variables worked synergistically, antagonistically, were additive, etc.). ANCOVA and MANCOVA...
Showing: 1 - 37 of 37 results