Fibronectin Recombinant Rabbit Monoclonal Antibody [PSH14-22]
cat.: HA723611
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse |
| Applications: | WB, IF-Cell, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH14-22 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage buffer: | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 272 kDa |
| Isotype: | IgG |
| Positive control: | HepG2 cell lysate, NIH/3T3 cell lysate, HepG2, NIH/3T3. |
| Subcellular location: | Secreted, extracellular space, extracellular matrix. |
| Recommended Dilutions:
WB IF-Cell FC IP |
1:5,000 1:3,000 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P02751 Human | P11276 Mouse |
| Alternative names: | CIG Cold insoluble globulin Cold-insoluble globulin DKFZp686F10164 DKFZp686H0342 DKFZp686I1370 DKFZp686O13149 ED B Fibronectin 1 FINC FINC_HUMAN FN FN1 FNZ GFND GFND2 LETS Migration stimulating factor MSF Ugl-Y3 |
Images
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Fig1:
Western blot analysis of Fibronectin on different lysates with Rabbit anti-Fibronectin antibody (HA723611) at 1/5,000 dilution. Lane 1: HepG2 cell lysate Lane 2: NIH/3T3 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 272 kDa Observed band size: 272 kDa Exposure time: 11 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723611) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Immunocytochemistry analysis of HepG2 cells labeling Fibronectin with Rabbit anti-Fibronectin antibody (HA723611) at 1/3,000 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Fibronectin antibody (HA723611) at 1/3,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Immunocytochemistry analysis of NIH/3T3 cells labeling Fibronectin with Rabbit anti-Fibronectin antibody (HA723611) at 1/3,000 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Fibronectin antibody (HA723611) at 1/3,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Flow cytometric analysis of HepG2 cells labeling Fibronectin. Cells were fixed and permeabilized. Then stained with the primary antibody (HA723611, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig5:
Flow cytometric analysis of NIH/3T3 cells labeling Fibronectin. Cells were fixed and permeabilized. Then stained with the primary antibody (HA723611, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig6:
Fibronectin was immunoprecipitated from 0.2 mg HepG2 cell lysate with HA723611 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723611 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HepG2 cell lysate (input) Lane 2: HA723611 IP in HepG2 cell lysate Lane 3: Rabbit IgG instead of HA723611 in HepG2 cell lysate Blocking/Dilution buffer: primary antibody dilution (K1803) Exposure time: 1 minute 34 seconds; ECL: K1801 |
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