- 710659 - Invitrogen Antibodies KLF4 antibody

No submitted references: Submit reference

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of HEK-293 (lane1), NCCIT (lane2), HT 29 (lane3), HCT 116 (lane4) and COLO 205 (lane 5). The blots were probed with Anti-KLF4 Recombinant Rabbit Polyclonal Antibody (Product # 710659, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A clear 45 kDa band corresponding to KLF4 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Expression of KLF4 was detected after transfection of HEK-293 cells with 50nM Silencer® Select KLF4 siRNA (Product # s17795, s17793) (lane 2, 3 respectively) and untransfected (lane 1) using KLF4 Recombinant Rabbit Polyclonal Antibody (Product # 710659, 1-2 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Relative density of the bands normalized to tubulin confirms silencing of KLF4 expression and the specificity of KLF4 Recombinant Rabbit Polyclonal Antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence was performed on fixed and permeabilized HEK 293 cells for detection of KLF4 using Anti-KLF4 Recombinant Rabbit Polyclonal Antibody (Product # 710659, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 0.4 µg/mL, 1:2500). Panel a) shows representative cells that were stained for detection and localization of KLF4 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938, 1:50). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 594 Phalloidin (Product # A12381, 1:200). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear localization of KLF4. Panel e) represents control cells with no primary Antibody to assess background.

710659