Fibrillarin Recombinant Rabbit Monoclonal Antibody [JE04-54]
cat.: HA722842
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | JE04-54 |
| 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*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 34 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within human Fibrillarin aa 272-321 / 321. |
| Positive control: | HeLa cell lysate, HepG2 cell lysate, O-2 OS cell lysate, NIH/3T3 cell lysate, Neuro-2a cell lysate, C6 cell lysate, PC-12 cell lysate, HeLa, NIH/3T3, PC-12. |
| Subcellular location: | Nucleus, nucleolus, Nucleus, nucleoplasm. |
| Recommended Dilutions:
WB IF-Cell FC IP |
1:1,000 1:1,000 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: P22087 Human | P35550 Mouse | P22509 Rat |
| Alternative names: | 34 kD nucleolar scleroderma antigen 34 kDa nucleolar scleroderma antigen Fbl FBRL_HUMAN FIB FIB1 FLRN Histone-glutamine methyltransferase Nop1p RNA U3 small nucleolar interacting protein 1 RNU3IP1 rRNA 2' O methyltransferase fibrillarin rRNA 2'-O-methyltransferase fibrillarin |
Images
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Fig1:
Western blot analysis of Fibrillarin on different lysates with Rabbit anti-Fibrillarin antibody (HA722842) at 1/1,000 dilution. Lane 1: HeLa cell lysate Lane 2: HepG2 cell lysate Lane 3: O-2 OS cell lysate Lane 4: NIH/3T3 cell lysate Lane 5: Neuro-2a cell lysate Lane 6: C6 cell lysate Lane 7: PC-12 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 34 kDa Observed band size: 37 kDa Exposure time: 20 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 (HA722842) at 1/1,000 dilution was used in 5% NFDM/TBST 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 HeLa cells labeling Fibrillarin with Rabbit anti-Fibrillarin antibody (HA722842) at 1/1,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Fibrillarin antibody (HA722842) at 1/1,000 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 (M1305-2, 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 Fibrillarin with Rabbit anti-Fibrillarin antibody (HA722842) at 1/1,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Fibrillarin antibody (HA722842) at 1/1,000 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 (M1305-2, 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:
Immunocytochemistry analysis of PC-12 cells labeling Fibrillarin with Rabbit anti-Fibrillarin antibody (HA722842) at 1/1,000 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Fibrillarin antibody (HA722842) at 1/1,000 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 (M1305-2, 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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Fig5:
Flow cytometric analysis of HeLa cells labeling Fibrillarin. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722842, 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:
Fibrillarin was immunoprecipitated from 0.2 mg HepG2 cell lysate with HA722842 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722842 at 1/1,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody (M1003-7) at 1/100,000 dilution was used for 1 hour at room temperature. Lane 1: HepG2 cell lysate (input) Lane 2: HA722842 IP in HepG2 cell lysate Lane 3: Rabbit IgG instead of HA722842 in HepG2 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 20 seconds; ECL: K1801 |
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