Anti-KLF4 antibody [ EPR3550(2)(ABC)]

• Supplier: Abcam
• Catalog: ab151733
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: EPR3550(2)(ABC)
• Applications: WB ICC/IF ICC/IF IHC-P
• Species Reactivities: Mouse, Rat, Human
• Antigen: Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human KLF4 aa 300-400
• Blocking Peptide: KLF4 peptide
• Description: Rabbit Monoclonal
• Gene: KLF4
• Conjugate: Unconjugated

Product images

All lanes : Anti-KLF4 antibody [ EPR3550(2)(ABC)] (ab151733) at 1/10000 dilution (purified)Lane 1 : HeLa cell lysateLane 2 : NCCIT cell lysateLane 3 : HEK293 cell lysateLysates/proteins at 20 µg per lane.SecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

All lanes : Anti-KLF4 antibody [ EPR3550(2)(ABC)] (ab151733) at 1/10000 dilution (purified)Lane 1 : RAW264.7 cell lysateLane 2 : PC-12 cell lysateLysates/proteins at 20 µg per lane.SecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

All lanes : Anti-KLF4 antibody [ EPR3550(2)(ABC)] (ab151733) at 1/1000 dilution (purified)Lane 1 : Mouse brain tissue lysateLane 2 : Mouse spleen tissue lysateLane 3 : C6 cell lysateLane 4 : NIH/3T3 cell lysateLysates/proteins at 20 µg per lane.SecondaryPeroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

All lanes : Anti-KLF4 antibody [ EPR3550(2)(ABC)] (ab151733) at 1/1000 dilution (unpurified)Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLane 3 : Caco 2 (Human colonic carcinoma cell line) Whole Cell LysateLane 4 : HUES7 (Human embryonic stem cell line) Whole Cell LysateLane 5 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/10000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

All lanes : Anti-KLF4 antibody [ EPR3550(2)(ABC)] (ab151733) at 1/1000 dilution (unpurified)Lane 1 : HeLa cell lysatesLane 2 : NCCIT cell lysatesLane 3 : 293T cell lysatesSecondaryGoat anti-rabbit HRP at 1/2000 dilution

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human squamous cell carcinoma of the cervix tissue labelling KLF4 with purified ab151733 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat stomach tissue labelling KLF4 with purified ab151733 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

IHC image of KLF4 staining in Human normal colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with unpurified ab151733, 1/100 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

IHC image of KLF4 staining in Mouse normal colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with unpurified ab151733, 1/100 dilution, for 15 mins at room temperature. A Goat anti-Rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling KLF4 with purified ab151733 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.Control: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

ICC/IF image of unpurified ab151733 stained Mouse embryonic stem cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab151733, 1/500 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Immunocytochemistry/Immunofluorescence analysis of human embryonic stem cells labelling KLF4 with unpurified ab151733 at 1/250. Cells were fixed with paraformaldehyde and then incubated the primary antibody for 1.5 hours at room temperature. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody (1/200).See Abreview