MA3-926

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Sodiµm/Calcium Exchanger using Anti-Sodiµm/Calcium Exchanger Monoclonal Antibody (C2C12) (Product # MA3-926) shows staining in A2058 Cells. Sodiµm/Calcium Exchanger staining (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 or an antibody recognizing Sodiµm/Calcium Exchanger (Product # MA3-926) at a dilution of 1:100 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503, Goat Anti-Mouse). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Sodiµm/Calcium Exchanger using Anti-Sodiµm/Calcium Exchanger Monoclonal Antibody (C2C12) (Product # MA3-926) shows staining in A549 Cells. Sodiµm/Calcium Exchanger staining (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 or an antibody recognizing Sodiµm/Calcium Exchanger (Product # MA3-926) at a dilution of 1:100 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503, Goat Anti-Mouse). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Sodium/Calcium Exchanger was performed using 70% confluent log phase SH-SY5Y cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Sodium/Calcium Exchanger Monoclonal Antibody (C2C12) (Product # MA3-926) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing predominantly plasma membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60x magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Sodiµm/Calcium Exchanger using Anti-Sodiµm/Calcium Exchanger Monoclonal Antibody (C2C12) (Product # MA3-926) shows staining in U251 Cells. Sodiµm/Calcium Exchanger staining (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 or an antibody recognizing Sodiµm/Calcium Exchanger (Product # MA3-926) at a dilution of 1:100 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503, Goat Anti-Mouse). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal deparaffinized Human heart tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:20 with a mouse monoclonal antibody recognizing Sodiµm/Calcium Exchanger (Product # MA3-926) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal deparaffinized Human kidney tissue tissues. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a mouse monoclonal antibody recognizing Sodiµm/Calcium Exchanger (Product # MA3-926) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3. Mass-tagging of cysteine substitution mutants in the cysteine-free NCX1 (C-). The indicated mutant NCX1 proteins were expressed in High Five cells and subjected to mass-tagging with the 5 kDa MALPEG reagent as described under Materials and Methods. Shown are western blots for the dNCX1 protein (C2C12 antibody) obtained from cell extracts. Left panels show untreated cells (lane 1) and cells treated with MALPEG and then sorted with FACS (lane 2); right panel represent cells treated with MALPEG in the presence of saponin. The arrow indicates the pegylated protein. As seen with mass-tagging of NCKX2, the observed mass shift always appeared greater than 5kDa, presumably due to the distinct physicochemical nature of PEG.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Molecular and cellular characterization of TiPSC-CMs. (A) Immunostaining of proteins involved in cardiac functionality in cardiomyocytes derived from T07. Scale bar shows 50 um. (B) Electron microscopy analysis of cardiomyocytes derived from T07. Upper micrograph shows sarcomere structures of cardiomyocytes derived from T07. White arrows indicate the Z lines of sarcomere structures. Scale bar shows 500 nm. Lower micrograph shows gap junctions in cardiomyocytes derived from T07 (white arrow). Scale bar shows 400 nm. (C) Line-scan image of Ca 2+ transient(lower figure) and its average fluorescence intensity in cardiomyocytes derived from T07 (upper graph) were shown. F/F 0 means fluorescence (F) normalized to baseline fluorescence (F 0 ).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 Nav1.5 protein expression is reduced in F at 10 -deficient ventricular myocytes. A Mass spectroscopic analysis was performed to detect protein expression in heart tissue samples from Fat10 fl/fl mice ( n = 3) and cFat10 -/- mice ( n = 3). Heatmap showing the differential expression of membrane proteins. B , C Representative immunoblots ( B ) and quantitative data ( C ) of membrane Nav1.5 protein expression in heart tissues from Fat10 fl/fl and cFat10 -/- mice (* p < 0.05); NCX: Na + /Ca 2+ exchanger. D , E Representative immunoblots ( D ) and quantitative data ( E ) of total Nav1.5 protein expression in heart extracts from cFat10 -/- and Fat10 fl/fl mice as detected by WB (* p < 0.05). F , G Representative immunoblots ( F ) and quantitative data ( G ) of cytoplasmic Nav1.5 expression in heart tissues from cFat10 - /- and Fat10 fl/fl mice (* p < 0.05) as detected by WB. H , I Representative immunostaining showing the degree of colocalization between Nav1.5 and Na + /K + ATPase ( H ) and Pearson's correlation coefficient for colocalization ( I ) in isolated cardiomyocytes (scale bar = 20 mum). All results are expressed as the mean +- SEM of independent experiments.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Cardiac myocyte Ca 2+ -handling proteins in mouse mammary tumor virus-polyomavirus middle T antigen (MMTV-PyMT)- and MMTV-PyMT+ mice--phospholamban ( A ); SERCA2a (sarcoplasmic or endoplasmic reticulum calcium 2 ATPase) ( B ); phosphorilated phospholamban (p-phospholamban) ( C ); sodium/calcium exchanger (NCX) ( D ); and SERCA2a/phospholambam ratio ( E ). Western blot bands are shown for each protein. In the MMTV-PyMT+ mice, phospholamban protein expression was increased ( P =0.011) and SERCA2a protein expression was not different between the MMTV-PyMT+ and MMTV-PyMT- groups ( P =0.165). SERCA2a/phospholamban ratio was reduced in the MMTV-PyMT+ mice ( P =0.007). p-Phospholamban and NCX protein expression were decreased ( P =0.038 and P =0.017, respectively) compared with MMTV-PyMT- mice. Mann-Whitney test. Values are expressed as medians and interquartile ranges (IQRs). Animals per group: 7 MMTV-PyMT- and 7 MMTV-PyMT+.