PA5-30992

antibody from Invitrogen Antibodies

Targeting: DHFR

Immunoprecipitation

Antibody data

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Validation data

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. Predicted reactivity: Mouse (88%), Rat (89%), Pig (88%), Bovine (85%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
Reactivity: Human, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 1.54 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

DHFR protein structure - PA5-30992 shown in red.

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.

Supportive validation

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.

Supportive validation

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.

Supportive validation

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.

Supportive validation

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.