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- Product number
- PA5-14896 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CYP3A4 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Vial size
- 200 µL
- Concentration
- 0.5 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
Submitted references β-Naphtoflavone and Ethanol Induce Cytochrome P450 and Protect towards MPP⁺ Toxicity in Human Neuroblastoma SH-SY5Y Cells.
Fernandez-Abascal J, Ripullone M, Valeri A, Leone C, Valoti M
International journal of molecular sciences 2018 Oct 28;19(11)
International journal of molecular sciences 2018 Oct 28;19(11)
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis using a CYP3A4 polyclonal antibody (Product # PA5-14896) in NCI-H460 cell lysates (35 µg per lane).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), SNU-5 (Lane 2), HT-29 (Lane 3), COLO 205 (Lane 4), A-431 (Lane 5), tissue extracts of Mouse Liver (Lane 6), Rat Liver (Lane 7), Mouse Brain (Lane 8), Rat Brain (Lane 9) and Mouse Kidney (Lane 10). The blot was probed with CD73 Polyclonal Antibody (Product # PA5-14896, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A band at 58 kDa corresponding to CYP3A4 was observed across the cell lines and tissues tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis in formalin-fixed, paraffin-embedded human liver tissue using a CYP3A4 polyclonal antibody (Product # PA5-14896), followed by HRP-conjugated secondary antibody and DAB staining.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of CEM cells using a CYP3A4 polyclonal antibody (Product # PA5-14896) (right) compared to a negative control cell (left) at a dilution of 1:10-50, followed by a FITC-conjugated goat anti-rabbit antibody
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 2 Effect of beta-NF and EtOH treatment on cytochrome P450 (CYP) isoforms' expression in undifferentiated SH-SY5Y cells. ( a - d ) The graphs show the relative protein levels quantification by Western blot for the CYP2D6 ( a ), 2E1 ( b ), 1A1 ( c ), and 3A4 ( d ) isoforms after treating cells with the inducers for 48 h (see Section 4 ). Data was acquired by measuring the fluorescent intensity per pixel on each band. The relative amount of protein normalised with beta-actin as a housekeeping protein for each condition was plotted as fold-increase and compared to the control, which was given a value of 1. Columns represent the mean +- SEM of at least three different experiments. Statistical significance was analyzed by one-way ANOVA followed by a Tukey post-test (* p < 0.05; ** p < 0.01). ( e ) Top panel shows a representative blot of beta-actin housekeeping protein detected with secondary antibody Cy3 (green) in control (2nd lane), beta-NF (3rd lane) and EtOH (4th lane) treatments. ( e ) Bottom panel shows representative blots of each isoform detected with secondary antibody Cy5 (red) in the mentioned conditions.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 2 Effect of beta-NF and EtOH treatment on cytochrome P450 (CYP) isoforms' expression in undifferentiated SH-SY5Y cells. ( a - d ) The graphs show the relative protein levels quantification by Western blot for the CYP2D6 ( a ), 2E1 ( b ), 1A1 ( c ), and 3A4 ( d ) isoforms after treating cells with the inducers for 48 h (see Section 4 ). Data was acquired by measuring the fluorescent intensity per pixel on each band. The relative amount of protein normalised with beta-actin as a housekeeping protein for each condition was plotted as fold-increase and compared to the control, which was given a value of 1. Columns represent the mean +- SEM of at least three different experiments. Statistical significance was analyzed by one-way ANOVA followed by a Tukey post-test (* p < 0.05; ** p < 0.01). ( e ) Top panel shows a representative blot of beta-actin housekeeping protein detected with secondary antibody Cy3 (green) in control (2nd lane), beta-NF (3rd lane) and EtOH (4th lane) treatments. ( e ) Bottom panel shows representative blots of each isoform detected with secondary antibody Cy5 (red) in the mentioned conditions.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 7 Immunostaining of CYP3A4 in SH-SY5Y cells after beta-NF and EtOH treatment. Representative immunofluorescence images of each experimental condition. Column 1 represents mitochondrial stain (red), column 2 represents ER-Green Fluorescent Protein (GFP) stain (green), and column 3 reports the CYP3A4 staining (blue). Column 4 represents the merge between CYP3A4 and mitochondria channel, column 5 represents the merge between CYP3A4 and ER-GFP, and column 6 represents the merge of the three channels. R Values: Pearson's correlation coefficient. Scale bar: 20 um.
