KLF4 Rabbit Polyclonal Antibody

• Catalog Number: orb15894
• Category: Antibodies
• Description: KLF4 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Equine, Gallus, Guinea pig, Porcine, Sheep
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human KLF4 (415-513/513aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:50-200, IHC-F=1:50-200, ICC/IF=1:100, IF=1:50-200, Flow-Cyt=1ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 56 kDa
• Target: KLF4
• UniProt ID: O43474
• RRID: AB_10767725
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Endothelial Kruppel like zinc finger protein; EZF;
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: K562. Primary Antibody (green line): Rabbit Anti-KLF4 antibody (orb15894), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Blank control: MCF7 (blue). Primary Antibody: Rabbit Anti-KLF4 AF647 Conjugated antibody, dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG/AF647 (orange), used under the same conditions. Protocol, The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with ice-cold 90% methanol for 30 min on ice. The cells were washed twice with 1X PBS. The cells were incubated in 1X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions followed by the incubated with antibody (1 µg/1x10^6 cells) for 30 min on ice. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (Rat lung), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (KLF4) Polyclonal Antibody, Unconjugated (orb15894) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Sample: 293T (human) cell Lysate at 40 ug, Primary: Anti-KLF4 (orb15894) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 56 kD, Observed band size: 60 kD.

Sample: Hela Cell (Human) Lysate at 30 ug, Primary: Anti-KLF4 (orb15894) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 56kD, Observed band size: 56kD.

Sample: Lane 1: Mouse NIH/3T3 cell lysates, Lane 2: Human A431 cell lysates, Lane 3: Human LOVO cell lysates, Lane 4: Human HeLa cell lysates, Lane 5: Human MCF-7 cell lysates, Lane 6: Human A549 cell lysates, Primary: Anti-KLF4 (orb15894) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 56 kDa, Observed band size: 50 kDa.