A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide bacteria and archaea with adaptive immunity against viruses and plasmids by using CRISPR RNAs (crRNAs) to guide the silencing of invading nucleic acids. We show here that in a subset of these systems, the mature crRNA that is base-paired to trans-activating crRNA (tracrRNA) forms a two-RNA structure that directs the CRISPR-associated protein Cas9 to introduce double-stranded (ds) breaks in target DNA.
At sites complementary to the crRNA-guide sequence, the Cas9 HNH nuclease domain cleaves the complementary strand, whereas the Cas9 RuvC-like domain cleaves the noncomplementary strand. The dual-tracrRNA:crRNA, when engineered as a single RNA chimera, also directs sequence-specific Cas9 dsDNA cleavage. Our study reveals a family of endonucleases that use dual-RNAs for site-specific DNA cleavage and highlights the potential to exploit the system for RNA-programmable genome editing.
Inherited genetic variation has a critical but as yet largely uncharacterized role in human disease. Here we report a public database of common variation in the human genome: more than one million single nucleotide polymorphisms (SNPs) for which accurate and complete genotypes have been obtained in 269 DNA samples from four populations, including ten 500-kilobase regions in which essentially all information about common DNA variation has been extracted.
These data document the generality of recombination hotspots, a block-like structure of linkage disequilibrium and low haplotype diversity, leading to substantial correlations of SNPs with many of their neighbours. We show how the HapMap resource can guide the design and analysis of genetic association studies, shed light on structural variation and recombination, and identify loci that may have been subject to natural selection during human evolution.
Identification and analysis of functional elements in 1% of the human genome by the ENCODE pilot project.
We report the generation and analysis of functional data from multiple, diverse experiments performed on a targeted 1% of the human genome as part of the pilot phase of the ENCODE Project. These data have been further integrated and augmented by a number of evolutionary and computational analyses. Together, our results advance the collective knowledge about human genome function in several major areas.
First, our studies provide convincing evidence that the genome is pervasively transcribed, such that the majority of its bases can be found in primary transcripts, including non-protein-coding transcripts, and those that extensively overlap one another. Second, systematic examination of transcriptional regulation has yielded new understanding about transcription start sites, including their relationship to specific regulatory sequences and features of chromatin accessibility and histone modification. Third, a more sophisticated view of chromatin structure has emerged, including its inter-relationship with DNA replication and transcriptional regulation.
Finally, integration of these new sources of information, in particular with respect to mammalian evolution based on inter- and intra-species sequence comparisons, has yielded new mechanistic and evolutionary insights concerning the functional landscape of the human genome. Together, these studies are defining a path for pursuit of a more comprehensive characterization of human genome function.
Rapid and efficient site-specific mutagenesis without phenotypic selection
Several single-base substitution mutations have been introduced into the lacZ alpha gene in cloning vector M13mp2, at 40-60% efficiency, in a rapid procedure requiring only transfection of the unfractionated products of standard in vitro mutagenesis reactions. Two simple additional treatments of the DNA, before transfection, produce a site-specific mutation frequency approaching 100%.
The approach is applicable to phenotypically silent mutations in addition to those that can be selected. The high efficiency, approximately equal to 10-fold greater than that observed using current methods without enrichment procedures, is obtained by using a DNA template containing several uracil residues in place of thymine. This template has normal coding potential for the in vitro reactions typical of site-directed mutagenesis protocols but is not biologically active upon transfection into a wild-type (i.e., ung+) Escherichia coli host cell.
Expression of the desired change, present in the newly synthesized non-uracil-containing covalently closed circular complementary strand, is thus strongly favored. The procedure has been applied to mutations introduced via both oligonucleotides and error-prone polymerization. In addition to its utility in changing DNA sequences, this approach can potentially be used to examine the biological consequences of specific lesions placed at defined positions within a gene.
Comprehensive molecular characterization of human colon and rectal cancer.
To characterize somatic alterations in colorectal carcinoma, we conducted a genome-scale analysis of 276 samples, analysing exome sequence, DNA copy number, promoter methylation and messenger RNA and microRNA expression. A subset of these samples (97) underwent low-depth-of-coverage whole-genome sequencing.
In total, 16% of colorectal carcinomas were found to be hypermutated: three-quarters of these had the expected high microsatellite instability, usually with hypermethylation and MLH1 silencing, and one-quarter had somatic mismatch-repair gene and polymerase ε (POLE) mutations. Excluding the hypermutated cancers, colon and rectum cancers were found to have considerably similar patterns of genomic alteration.
Twenty-four genes were significantly mutated, and in addition to the expected APC, TP53, SMAD4, PIK3CA and KRAS mutations, we found frequent mutations in ARID1A, SOX9 and FAM123B. Recurrent copy-number alterations include potentially drug-targetable amplifications of ERBB2 and newly discovered amplification of IGF2. Recurrent chromosomal translocations include the fusion of NAV2 and WNT pathway member TCF7L1. Integrative analyses suggest new markers for aggressive colorectal carcinoma and an important role for MYC-directed transcriptional activation and repression.
SPAG1 Conjugated Antibody |
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C31270 | SAB | 100ul | EUR 397 |
SPAG1 siRNA |
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20-abx934789 | Abbexa |
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SPAG1 siRNA |
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20-abx934790 | Abbexa |
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SPAG1 siRNA |
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20-abx905218 | Abbexa |
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SPAG1 Rabbit pAb |
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A3087-100ul | Abclonal | 100 ul | EUR 308 |
SPAG1 Rabbit pAb |
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A3087-200ul | Abclonal | 200 ul | EUR 459 |
SPAG1 Rabbit pAb |
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A3087-20ul | Abclonal | 20 ul | Ask for price |
SPAG1 Rabbit pAb |
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A3087-50ul | Abclonal | 50 ul | EUR 223 |
Polyclonal Goat anti-GST α-form |
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GST-ANTI-1 | Detroit R&D | 50 uL | EUR 280 |
Polyclonal Goat anti-GST μ-form |
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GST-ANTI-2 | Detroit R&D | 50 uL | EUR 280 |
Polyclonal Goat anti-GST p-form |
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GST-ANTI-3 | Detroit R&D | 50 uL | EUR 280 |
Mouse SPAG1 shRNA Plasmid |
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20-abx973799 | Abbexa |
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Rat SPAG1 shRNA Plasmid |
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20-abx989852 | Abbexa |
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Human SPAG1 shRNA Plasmid |
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20-abx954551 | Abbexa |
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Human SPAG1 ELISA Kit |
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ELA-E9879h | Lifescience Market | 96 Tests | EUR 824 |
SPAG1 ELISA KIT|Human |
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EF006557 | Lifescience Market | 96 Tests | EUR 689 |
Sperm Associated Antigen 1 (SPAG1) Antibody |
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20-abx131166 | Abbexa |
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Sperm-Associated Antigen 1 (SPAG1) Antibody |
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20-abx007339 | Abbexa |
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Sperm Associated Antigen 1 (SPAG1) Antibody |
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20-abx339805 | Abbexa |
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Sperm Associated Antigen 1 (SPAG1) Antibody |
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20-abx339980 | Abbexa |
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SPAG1 ELISA Kit (Mouse) (OKEH04948) |
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OKEH04948 | Aviva Systems Biology | 96 Wells | EUR 779 |
Description: Description of target: May play a role in the cytoplasmic assembly of the ciliary dynein arms. May play a role in fertilization. Binds GTP and has GTPase activity.;Species reactivity: Mouse;Application: ;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 0.079 ng/mL |
SPAG1 ELISA Kit (Human) (OKEH02027) |
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OKEH02027 | Aviva Systems Biology | 96 Wells | EUR 779 |
Description: Description of target: The correlation of anti-sperm antibodies with cases of unexplained infertility implicates a role for these antibodies in blocking fertilization. Improved diagnosis and treatment of immunologic infertility, as well as identification of proteins for targeted contraception, are dependent on the identification and characterization of relevant sperm antigens. The protein expressed by this gene is recognized by anti-sperm agglutinating antibodies from an infertile woman. Furthermore, immunization of female rats with the recombinant human protein reduced fertility. This protein localizes to the plasma membrane of germ cells in the testis and to the post-acrosomal plasma membrane of mature spermatozoa. Recombinant polypeptide binds GTP and exhibits GTPase activity. Thus, this protein may regulate GTP signal transduction pathways involved in spermatogenesis and fertilization. Two transcript variants of this gene encode the same protein.;Species reactivity: Human;Application: ELISA;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 0.023 ng/mL |
SPAG1 ELISA Kit (Rat) (OKEH05931) |
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OKEH05931 | Aviva Systems Biology | 96 Wells | EUR 779 |
Description: Description of target: May play a role in the cytoplasmic assembly of the ciliary dynein arms. Binds GTP and has GTPase activity. Plays a role in fertilization.;Species reactivity: Rat;Application: ;Assay info: Assay Methodology: Quantitative Sandwich ELISA;Sensitivity: 0.08 ng/mL |
Spag1 ORF Vector (Mouse) (pORF) |
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ORF058232 | ABM | 1.0 ug DNA | EUR 506 |
SPAG1 ORF Vector (Human) (pORF) |
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ORF033209 | ABM | 1.0 ug DNA | EUR 405 |
Spag1 ORF Vector (Rat) (pORF) |
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ORF076867 | ABM | 1.0 ug DNA | EUR 506 |
Recombinant Sperm Associated Antigen 1 (SPAG1) |
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4-RPC865Hu01 | Cloud-Clone |
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Description: Recombinant Human Sperm Associated Antigen 1 expressed in: E.coli |
SPAG1 sgRNA CRISPR Lentivector set (Human) |
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K2264001 | ABM | 3 x 1.0 ug | EUR 339 |
Spag1 sgRNA CRISPR Lentivector set (Rat) |
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K6315201 | ABM | 3 x 1.0 ug | EUR 339 |
Sperm Associated Antigen 1 (SPAG1) Polyclonal Antibody (Human, Rat) |
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4-PAC865Hu01 | Cloud-Clone |
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Description: A Rabbit polyclonal antibody against Human, Rat Sperm Associated Antigen 1 (SPAG1) |
Human Sperm Associated Antigen 1 (SPAG1) Protein |
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20-abx650220 | Abbexa |
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SPAG1 sgRNA CRISPR Lentivector (Human) (Target 1) |
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K2264002 | ABM | 1.0 ug DNA | EUR 154 |
SPAG1 sgRNA CRISPR Lentivector (Human) (Target 2) |
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K2264003 | ABM | 1.0 ug DNA | EUR 154 |
SPAG1 sgRNA CRISPR Lentivector (Human) (Target 3) |
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K2264004 | ABM | 1.0 ug DNA | EUR 154 |
Spag1 sgRNA CRISPR Lentivector (Rat) (Target 1) |
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K6315202 | ABM | 1.0 ug DNA | EUR 154 |
Spag1 sgRNA CRISPR Lentivector (Rat) (Target 2) |
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K6315203 | ABM | 1.0 ug DNA | EUR 154 |
Spag1 sgRNA CRISPR Lentivector (Rat) (Target 3) |
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K6315204 | ABM | 1.0 ug DNA | EUR 154 |
Spag1 3'UTR Luciferase Stable Cell Line |
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TU221015 | ABM | 1.0 ml | Ask for price |
Spag1 3'UTR GFP Stable Cell Line |
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TU271015 | ABM | 1.0 ml | Ask for price |
SPAG1 3'UTR Luciferase Stable Cell Line |
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TU024373 | ABM | 1.0 ml | EUR 1394 |
SPAG1 3'UTR GFP Stable Cell Line |
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TU074373 | ABM | 1.0 ml | EUR 1394 |
SPAG1 Protein Vector (Mouse) (pPB-C-His) |
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PV232926 | ABM | 500 ng | EUR 1065 |
SPAG1 Protein Vector (Mouse) (pPB-N-His) |
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PV232927 | ABM | 500 ng | EUR 1065 |
SPAG1 Protein Vector (Mouse) (pPM-C-HA) |
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PV232928 | ABM | 500 ng | EUR 1065 |
SPAG1 Protein Vector (Mouse) (pPM-C-His) |
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PV232929 | ABM | 500 ng | EUR 1065 |
SPAG1 Protein Vector (Rat) (pPB-C-His) |
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PV307466 | ABM | 500 ng | EUR 1166 |
SPAG1 Protein Vector (Rat) (pPB-N-His) |
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PV307467 | ABM | 500 ng | EUR 1166 |
SPAG1 Protein Vector (Rat) (pPM-C-HA) |
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PV307468 | ABM | 500 ng | EUR 1166 |
SPAG1 Protein Vector (Rat) (pPM-C-His) |
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PV307469 | ABM | 500 ng | EUR 1166 |
SPAG1 Protein Vector (Human) (pPB-C-His) |
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PV132834 | ABM | 500 ng | Ask for price |
SPAG1 Protein Vector (Human) (pPB-N-His) |
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PV132835 | ABM | 500 ng | Ask for price |
SPAG1 Protein Vector (Human) (pPM-C-HA) |
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PV132836 | ABM | 500 ng | Ask for price |
SPAG1 Protein Vector (Human) (pPM-C-His) |
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PV132837 | ABM | 500 ng | Ask for price |
Sperm Associated Antigen 1 (SPAG1) Polyclonal Antibody (Human, Rat), APC |
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4-PAC865Hu01-APC | Cloud-Clone |
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Description: A Rabbit polyclonal antibody against Human, Rat Sperm Associated Antigen 1 (SPAG1). This antibody is labeled with APC. |
Sperm Associated Antigen 1 (SPAG1) Polyclonal Antibody (Human, Rat), Biotinylated |
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4-PAC865Hu01-Biotin | Cloud-Clone |
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Description: A Rabbit polyclonal antibody against Human, Rat Sperm Associated Antigen 1 (SPAG1). This antibody is labeled with Biotin. |
Sperm Associated Antigen 1 (SPAG1) Polyclonal Antibody (Human, Rat), FITC |
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4-PAC865Hu01-FITC | Cloud-Clone |
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Description: A Rabbit polyclonal antibody against Human, Rat Sperm Associated Antigen 1 (SPAG1). This antibody is labeled with FITC. |
Sperm Associated Antigen 1 (SPAG1) Polyclonal Antibody (Human, Rat), HRP |
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4-PAC865Hu01-HRP | Cloud-Clone |
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Description: A Rabbit polyclonal antibody against Human, Rat Sperm Associated Antigen 1 (SPAG1). This antibody is labeled with HRP. |
Sperm Associated Antigen 1 (SPAG1) Polyclonal Antibody (Human, Rat), PE |
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4-PAC865Hu01-PE | Cloud-Clone |
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Description: A Rabbit polyclonal antibody against Human, Rat Sperm Associated Antigen 1 (SPAG1). This antibody is labeled with PE. |
Sperm Associated Antigen 1 (SPAG1) Polyclonal Antibody (Human, Rat), Cy3 |
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4-PAC865Hu01-Cy3 | Cloud-Clone |
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Description: A Rabbit polyclonal antibody against Human, Rat Sperm Associated Antigen 1 (SPAG1). This antibody is labeled with Cy3. |
Mouse Sperm-associated antigen 1 (SPAG1) ELISA Kit |
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abx521376-96tests | Abbexa | 96 tests | EUR 739 |
Rat Sperm-associated antigen 1 (SPAG1) ELISA Kit |
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abx521377-96tests | Abbexa | 96 tests | EUR 739 |
Mouse Spag1/ Sperm-associated antigen 1 ELISA Kit |
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E1398Mo | Sunlong | 1 Kit | EUR 632 |
Rat Spag1/ Sperm-associated antigen 1 ELISA Kit |
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E0935Ra | Sunlong | 1 Kit | EUR 646 |
Human SPAG1/ Sperm-associated antigen 1 ELISA Kit |
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E2372Hu | Sunlong | 1 Kit | EUR 605 |
Human Sperm-associated antigen 1 (SPAG1) ELISA Kit |
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abx251897-96tests | Abbexa | 96 tests | EUR 739 |
Human SPAG1(Sperm-associated antigen 1) ELISA Kit |
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EH2525 | FN Test | 96T | EUR 567.6 |
Description: Method of detection: Double Antibody, Sandwich ELISA;Reacts with: Homo sapiens;Sensitivity: 0.094 ng/ml |
Human Sperm- associated antigen 1, SPAG1 ELISA KIT |
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ELI-40946h | Lifescience Market | 96 Tests | EUR 824 |
Mouse Sperm- associated antigen 1, Spag1 ELISA KIT |
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ELI-40947m | Lifescience Market | 96 Tests | EUR 865 |
SPAG1 Lentiviral Vector (Rat) (CMV) (pLenti-GIII-CMV) |
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LV660379 | ABM | 1.0 ug DNA | EUR 1355 |
SPAG1 Lentiviral Vector (Rat) (UbC) (pLenti-GIII-UbC) |
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LV660383 | ABM | 1.0 ug DNA | EUR 1355 |
SPAG1 Lentiviral Vector (Rat) (EF1a) (pLenti-GIII-EF1a) |
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LV660384 | ABM | 1.0 ug DNA | EUR 1355 |
Sperm Associated Antigen 1 (SPAG1) Polyclonal Antibody (Human, Rat), APC-Cy7 |
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4-PAC865Hu01-APC-Cy7 | Cloud-Clone |
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Description: A Rabbit polyclonal antibody against Human, Rat Sperm Associated Antigen 1 (SPAG1). This antibody is labeled with APC-Cy7. |
SPAG1 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Human) |
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K2264005 | ABM | 3 x 1.0 ug | EUR 376 |
Spag1 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Rat) |
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K6315205 | ABM | 3 x 1.0 ug | EUR 376 |
SPAG1 sgRNA CRISPR/Cas9 All-in-One Lentivector (Human) (Target 1) |
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K2264006 | ABM | 1.0 ug DNA | EUR 167 |
SPAG1 sgRNA CRISPR/Cas9 All-in-One Lentivector (Human) (Target 2) |
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K2264007 | ABM | 1.0 ug DNA | EUR 167 |
SPAG1 sgRNA CRISPR/Cas9 All-in-One Lentivector (Human) (Target 3) |
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K2264008 | ABM | 1.0 ug DNA | EUR 167 |
SPAG1 Lentiviral Vector (Rat) (CMV) (pLenti-GIII-CMV-C-term-HA) |
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LV660380 | ABM | 1.0 ug DNA | EUR 1355 |
SPAG1 Lentiviral Vector (Rat) (CMV) (pLenti-GIII-CMV-GFP-2A-Puro) |
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LV660381 | ABM | 1.0 ug DNA | EUR 1413 |
SPAG1 Lentiviral Vector (Rat) (CMV) (pLenti-GIII-CMV-RFP-2A-Puro) |
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LV660382 | ABM | 1.0 ug DNA | EUR 1413 |
Spag1 sgRNA CRISPR/Cas9 All-in-One Lentivector (Rat) (Target 1) |
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K6315206 | ABM | 1.0 ug DNA | EUR 167 |
Spag1 sgRNA CRISPR/Cas9 All-in-One Lentivector (Rat) (Target 2) |
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K6315207 | ABM | 1.0 ug DNA | EUR 167 |
Spag1 sgRNA CRISPR/Cas9 All-in-One Lentivector (Rat) (Target 3) |
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K6315208 | ABM | 1.0 ug DNA | EUR 167 |
Anti-Anti-SEPT6 antibody antibody |
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STJ11100949 | St John's Laboratory | 100 µl | EUR 277 |
Description: This gene is a member of the septin family of GTPases. Members of this family are required for cytokinesis. One version of pediatric acute myeloid leukemia is the result of a reciprocal translocation between chromosomes 11 and X, with the breakpoint associated with the genes encoding the mixed-lineage leukemia and septin 2 proteins. This gene encodes four transcript variants encoding three distinct isoforms. An additional transcript variant has been identified, but its biological validity has not been determined. |
Anti-Anti-SEPT9 Antibody antibody |
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STJ111369 | St John's Laboratory | 100 µl | EUR 277 |
Description: This gene is a member of the septin family involved in cytokinesis and cell cycle control. This gene is a candidate for the ovarian tumor suppressor gene. Mutations in this gene cause hereditary neuralgic amyotrophy, also known as neuritis with brachial predilection. A chromosomal translocation involving this gene on chromosome 17 and the MLL gene on chromosome 11 results in acute myelomonocytic leukemia. Multiple alternatively spliced transcript variants encoding different isoforms have been described. |
Anti-Anti-SEPT11 Antibody antibody |
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STJ111530 | St John's Laboratory | 100 µl | EUR 277 |
Anti-Anti-SEPT4 Antibody antibody |
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STJ112276 | St John's Laboratory | 100 µl | EUR 277 |
Description: This gene is a member of the septin family of nucleotide binding proteins, originally described in yeast as cell division cycle regulatory proteins. Septins are highly conserved in yeast, Drosophila, and mouse, and appear to regulate cytoskeletal organization. Disruption of septin function disturbs cytokinesis and results in large multinucleate or polyploid cells. This gene is highly expressed in brain and heart. Alternatively spliced transcript variants encoding different isoforms have been described for this gene. One of the isoforms (known as ARTS) is distinct; it is localized to the mitochondria, and has a role in apoptosis and cancer. |
Anti-Anti-MARCH9 Antibody antibody |
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STJ112609 | St John's Laboratory | 100 µl | EUR 277 |
Anti-Anti-SEPT2 Antibody antibody |
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STJ25475 | St John's Laboratory | 100 µl | EUR 277 |
Anti-Anti-SEPT5 Antibody antibody |
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STJ25477 | St John's Laboratory | 100 µl | EUR 277 |
Description: This gene is a member of the septin gene family of nucleotide binding proteins, originally described in yeast as cell division cycle regulatory proteins. Septins are highly conserved in yeast, Drosophila, and mouse and appear to regulate cytoskeletal organization. Disruption of septin function disturbs cytokinesis and results in large multinucleate or polyploid cells. This gene is mapped to 22q11, the region frequently deleted in DiGeorge and velocardiofacial syndromes. A translocation involving the MLL gene and this gene has also been reported in patients with acute myeloid leukemia. Alternative splicing results in multiple transcript variants. The presence of a non-consensus polyA signal (AACAAT) in this gene also results in read-through transcription into the downstream neighboring gene (GP1BB; platelet glycoprotein Ib), whereby larger, non-coding transcripts are produced. |
Anti-Anti-SEPT8 Antibody antibody |
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STJ25479 | St John's Laboratory | 100 µl | EUR 277 |
Description: This gene is a member of the septin family of nucleotide binding proteins, originally described in yeast as cell division cycle regulatory proteins. Septins are highly conserved in yeast, Drosophila, and mouse, and appear to regulate cytoskeletal organization. Disruption of septin function disturbs cytokinesis and results in large multinucleate or polyploid cells. Multiple alternatively spliced transcript variants encoding different isoforms have been found for this gene. |
Anti-Anti-SEPT2 Antibody antibody |
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STJ28365 | St John's Laboratory | 100 µl | EUR 277 |