MA1-745

antibody from Invitrogen Antibodies

Targeting: SNTB2 D16S2531E, EST25263, SNT2B2, SNT3, SNTL

Provider product page for MA1-745

Western blot

Immunocytochemistry

Immunoprecipitation

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [31]
Comments [0]
Validations
Western blot [3]
Immunocytochemistry [4]
Other assay [12]

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Product number: MA1-745 - Provider product page
Provider: Invitrogen Antibodies
Product name: SNTB2 Monoclonal Antibody (1351)
Antibody type: Monoclonal
Antigen: Purifed from natural sources
Description: MA1-745 detects alpha 1, beta 1 and beta 2 subunits of syntrophin. This antibody has been shown to react with human, mouse, rat, chicken, canine and amphibian samples. MA1-745 has been successfully used in Western blot, immunoprecipitation and immunofluorescence procedures. By Western blot, this antibody detects an ~58 kDa protein representing syntrophin from rat and mouse cell extracts. The MA1-745 immunogen is whole purified syntrophin from Torpedo californica electric organ postsynaptic membrane. Characterization studies of this antibody have demonstrated that clone 1351 is directed against an epitope within the PDZ domain of syntrophin. MA1-745, clone 1351, is seen as the "gold standard" for syntrophin assessment.
Reactivity: Human, Mouse, Rat, Canine, Chicken/Avian
Host: Mouse
Isotype: IgG
Antibody clone number: 1351
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

SNTB2 protein structure - MA1-745 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of U-87MG (Lane 1), PC 3 (Lane 2) and HeLa (Lane 3). The blots were probed with Anti-Syntrophin Mouse Monoclonal Antibody (Product # MA1-745, 0.1-1 µg/mL) and detected by chemiluminescence Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 62-6520, 1:4000 dilution). A 60 kDa band corresponding to Syntrophin was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System (Product # IB21001).The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of Syntrophin was achieved by transfecting U87MG with Syntrophin specific siRNA (Silencer® select Product # s13261 and s13263). Western blot analysis was performed using whole cell lysates from the Syntrophin knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-Syntrophin Mouse Monoclonal Antibody (Product # MA1-745, 0.1-1 µg/mL) and Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 62-6520, 1:4000 dilution). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to Syntrophin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Syntrophin was performed using Syntrophin Monoclonal Antibody (Product # MA1-745) on canine lysate.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of Syntrophin using Syntrophin Monoclonal Antibody (1351) (Product # MA1-745) shows staining in 293Cells. Syntrophin (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing Syntrophin (Product # MA1-745) at a dilution of 1:20 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of Syntrophin using Syntrophin Monoclonal Antibody (1351) (Product # MA1-745) shows staining in A2058 Cells. Syntrophin (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing Syntrophin (Product # MA1-745) at a dilution of 1:20 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Syntrophin using Syntrophin Monoclonal Antibody (1351) (Product # MA1-745) shows staining in Hela Cells. Syntrophin (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing Syntrophin (Product # MA1-745) at a dilution of 1:20 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of Syntrophin was done on 70% confluent log phase PC-3 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Syntrophin (1351) Mouse Monoclonal Antibody (Product # MA1-745) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

Supportive validation

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 10 WT/mdx muscle displays nNOS and dystrobrevin-alpha. Protein extracts from diaphragms of WT (lane 1), 10-30% WT/mdx (lane 2) and mdx (lane 3) mice were subjected to a western blot for dystrophin, nNOS, dystrobrevin-alpha, syntrophin and tubulin. *: dystrobrevin-alpha isoforms enriched in mdx and WT/mdx but not in WT muscle. Experiments were performed with pectoralis, diaphragm and quadriceps muscles of 3 animals (n = 3) per group (WT, 10-30% WT/mdx and mdx).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 11 Syntrophin is present in WT/mdx but not in the mdx muscle. a-f, diaphragms from WT (a, b), mdx (c, d), and 10-30% WT/mdx (e-f) mice were sectioned and stained for dystrophin (a, c, e) and syntrophin (b, d, f). Magnification: 200x. Experiments were performed with diaphragms of 2 animals (n = 2) per group (WT, mdx and 10-30% WT/mdx).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 iPSCs incorporate globally. a) PCR analysis showing that eGFP marker (indicative of iPSCs) is present globally in the WT/mdx chimera; b) Semi Q genomic PCR using DNA from quadriceps muscle. Note that the intensity of the Dys+ band (indicative of percentage of iPSC chimerism) in WT/mdx and WT/mdx:utrophin is approximately 1/5 of that observed in WT; c) WB analysis showing dystrophin, utrophin, dystrobrevin and syntrophin production in the quadriceps. Abbreviations: eGFP: enhanced green fluorescent protein, F: fat, Li: liver, Lu: lung, S: spleen, M: quadriceps muscle, T: tail, dys+: WT allele of dystrophin, Id+: WT allele of Id1 control (Id1 is present in both iPSC-derived and blastocyst-derived cells, and thus, is invariant among WT, KO and chimeric groups), Dys: 420 Kd dystrophin, Utr: 420 Kd utrophin, Dbv: 40 Kd (isoform of the dystrophin-dependent) dystrobrevin-alpha, Synt: 55 Kd syntrophin; Tub: 55 Kd tubulin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 The effect of 5-fluorouracil (5FU) and 5FU with BGP-15 (5FU+BGP) treatment on expression of cytoskeletal structural proteins. Western blotting experiments were undertaken in tibialis anterior (TA) muscle homogenate, with samples probed for ( A , B ) cytoskeletal structural proteins including; laminin (LAM), -dystroglycan (-DGC), -sarcoglycan (-SGC), dystrophin (DYS), dystrobrevin (DBVN), syntrophin (SYTN), desmin (DES) and talin (TAL). ( C ) Phosphorylated (Ser473) and total Akt were probed for as an indicator of mammalian target of rapamycin Complex 2 (mTORC2) activity. ( D ) Ankrd2 was probed for as a marker of mechano-sensitvit. ( E ) Representative images for phosphorylated Akt Ser473 , total Akt and Ankrd2 displayed. Protein expression was normalised to total protein derived from Coomassie Brilliant Blue (CBB) staining and presented relative to vehicle (VEH) control group. * = p < 0.05; n = 4 for VEH and n = 6-8 for 5FU and 5FU+BGP groups for Western blotting. Data are mean +- SEM.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Generation and characterization of alpha-syntrophin mice. A, A targeting vector was constructed using a 7.4-kb Not I (N)/XbaI (X) restriction fragment long arm and a 1.5-kb XbaI fragment short arm. Homologous recombination resulted in the deletion of 2.8-kb of the alpha-syntrophin gene including all of exon 1. B, Southern blot analysis was performed using genomic DNA (isolated from ES cells or from mice) digested with EcoRI (E). Hybridization using the 500-bp probe shown in A, detected a 6.2-kb wild-type band and a 10-kb recombinant band. C, RNA blot analysis of poly A + RNA from the mice indicated. The positions of RNA standards are indicated. D, Immunoblot of muscle proteins partially purified with mAb 1351 and detected with alpha-syntrophin and beta1-syntrophin isoform specific antibodies. The position of a 59-kD protein standard is indicated on the left.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Subcellular distribution of syntrophin and utrophin in INS-1 cells. Post-nuclear supernatants of INS-1 cells were fractionated on continuous sucrose density gradients (0.4-2 M). Aliquots of each gradient fraction were immunoblotted with antibodies directed against ICA512 (mICA512cyto), carboxypeptidase E, syntrophin (1351) and utrophin.