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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [5]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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- Product number
- PA5-30992 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DHFR Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, PC-12.
- Concentration
- 1.54 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of DHFR using 30 µg of A) 293T and B) A431 lysate. Samples were loaded onto a 12% SDS-PAGE gel and probed with a DHFR polyclonal antibody (Product # PA5-30992) at a dilution of 1:1000.
- 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), HEK-293 (Lane 2) and K-562 (Lane 3). The blot was probed with DHFR Polyclonal Antibody (Product # PA5-30992, 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 20kDa corresponding to DHFR was observed across the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using DHFR Polyclonal Antibody (Product # PA5-30992). Whole cell extract (30 µg) was separated by 12% SDS-PAGE, and the membrane was blotted with DHFR Polyclonal Antibody (Product # PA5-30992) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- DHFR Polyclonal Antibody detects DHFR protein by western blot analysis. A. 30 µg PC-12 whole cell lysate/extract.12% SDS-PAGE. DHFR Polyclonal Antibody (Product # PA5-30992) dilution: 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of DHFR was achieved by transfecting HEK-293 cells with DHFR specific siRNAs (Silencer® select Product # s4081, s4082). Western blot analysis (Fig. a) was performed using whole cell extracts from the DHFR knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with DHFR Polyclonal Antibody (Product # PA5-30992, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to DHFR.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of DHFR in paraformaldehyde-fixed A431 cells using a DHFR polyclonal antibody (Product # PA5-30992) at a 1:200 dilution.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using DHFR (Product # PA5-30992) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
