Rabbit Anti Trpv5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Autosomal Dominant Hypercalciuria in a Mouse Model Due to a Mutation of the Epithelial Calcium Channel, TRPV5"
Article Title: Autosomal Dominant Hypercalciuria in a Mouse Model Due to a Mutation of the Epithelial Calcium Channel, TRPV5
Journal: PLoS ONE
Figure Legend Snippet: Channel characteristics of wild-type and mutant TRPV5. ( A ) Whole-cell currents in TRPV5-WT (V5-WT) and TRPV5-682P (V5-S682P) injected Xenopus oocytes recorded in response to 300 ms test pulses to various potentials (from −100 to +60 mV in 10 mV increments). Holding potential, 0 mV (N = 5). ( B ) Mean current-voltage relationships for TRPV5-WT and TRPV5-682P channels (N = 5). These current-voltage relationships are similar to those reported for TRPV5 channels  . ( C ) Mean whole-cell tail currents measured in TRPV5-WT and TRPV5-682P injected Xenopus oocytes during test potentials applied in 10 mV increments from −70 to +40 mV after a pre-pulse to −100 mV in TRPV5-WT and TRPV5-682P channels (N = 5). ( D ) Time-dependent inhibition of TRPV-WT and TRPV5-682P whole-cell currents. Oocytes were stimulated every 1 s. The peak current amplitude was normalised to that recorded during the first pulse (N = 4). ( E ) Representative trace of Fura-2 ratio in HEK293 cells transiently transfected with an empty EGFP vector (mock), or EGFP-tagged TRPV5-WT or TRPV5-S682P. Cells expressing EGFP were selected and monitored for changes in intracellular Ca 2+ levels when extracellular Ca 2+ concentrations were varied from 1.4 mM Ca 2+ to 0 mM Ca 2+ (2 mM EDTA) and 1.4 mM Ca 2+ which was facilitated by superfusion. ( F ) Fura-2 levels under resting conditions (t0), minimal Fura-2 ratio after EDTA treatment (tmin) and peak level (tmax) upon administration of 1.4 mM Ca 2+ after EDTA treatment. Average data of cells transfected with the empty vector (n = 7), TRPV5-wt (n = 24) and TRPV5-S682P (N = 24) from at least three independent experiments. * p
Techniques Used: Mutagenesis, Injection, Mass Spectrometry, Inhibition, Transfection, Plasmid Preparation, Expressing
Figure Legend Snippet: Assessment of Renal Expression of Calcium Regulatory Genes and Proteins. Renal expression of ( A ) Trpv5 , ( B ) Trpv6 and ( C ) Cyp24a1 in wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice (n = 6/group) were assessed by quantitative real-time PCR. All data were normalised to levels of the housekeeping gene Gapdh and wild-type values are expressed as 1. Histogram data are presented as mean ± SEM. # p
Techniques Used: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction
Figure Legend Snippet: Histological and immunohistochemical assessment of kidneys from HCALC1 mice. Representative images in Trpv5 +/+ (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice of: ( A ) Masson's trichrome staining of renal cortex showing areas of interstitial fibrosis in Trpv5 682P/+ and Trpv5 682P/682P mice (light blue), ( B ) anti-CD3-labelling (green) showing a large number of T-lymphocytes present in the interstitial regions of the Trpv5 682P/+ and Trpv5 682P/682P mouse kidneys, ( C ) TUNEL-labelling (green) of the renal cortex showing the presence of tubular cell apoptosis in the Trpv5 682P/+ and Trpv5 682P/682P mouse kidneys. Scale bar = 50 µm. ( D ) Immunohistochemical images of kidney sections from wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice, co-stained for TRPV5 (green) and NCC (red). * denotes co-localisation. Scale bar = 50 µm. ( E ) Kidney sections from wild-type (wt), Trpv5 682P/+ (het) and Trpv5 682P/682P (hom) mice, co-stained for TRPV5 (green) and AQP2 (red). * denotes co-localisation. Scale bar = 50 µm.
Techniques Used: Immunohistochemistry, Mouse Assay, Staining, TUNEL Assay
Figure Legend Snippet: Hypercalciuria in HCALC1 ENU mutant mice and identification of a Trpv5 mutation. ( A ) Urine calcium/creatinine ratios in 23 G2 offspring of the HCALC1 founder male revealed that 10 of the 23 mice were hypercalciuric, consistent with an autosomal dominant inheritance. Bar, mean calcium/creatinine values. ( B ) Haplotype analysis of 89 G2 mice (39 hypercalciuric and 50 normocalciuric) was initially undertaken separately in the hypercalciuric and normocalciuric mice, as the penetrance of HCALC1 was unknown. Haplotype analysis of the hypercalciuric mice localised Hcalc1 to a 17.38 Mb interval on chromosome 6, flanked by rs13478688 and rs30110406 (broken double-headed arrow). Haplotype analysis using combined data for the hypercalciuric and normocalciuric mice identified the smaller interval, 11.94-Mb, flanked by rs13478709 and rs30110406 (solid double-headed arrow). The Hcalc1 locus is inherited with the C57BL/6J haplotype from the F1 founder male. Filled box, C57BL/6J allele; and open box, C3H/HeH allele. Number of mice observed for each haplotype is shown beneath each column. ( C ) DNA sequence analysis of Trpv5 identified a heterozygous T to C transition in codon 682 in hypercalciuric mice predicted to alter a wild-type serine (Ser) to a mutant proline (Pro). This mutation resulted in gain of a Bsa JI restriction enzyme site that was used to confirm the presence of the mutation in the 39 hypercalciuric mice (n = 3 shown) and its absence in the 50 normocalciuric mice (n = 3 shown). wt, wild-type; m, mutant. ( D ) Amino acid sequence alignment revealed evolutionary conservation of the wild-type mouse TRPV5 serine (S) residue at codon 682 (arrowed) in 5 species, as well as in mouse TRPV6 (mTrpv6). Identical residues are shaded black and conservative changes are shaded grey.
Techniques Used: Mutagenesis, Mouse Assay, Sequencing