Nucleolin Recombinant Rabbit Monoclonal Antibody [JE65-71]
cat.: HA721073
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IHC-P |
| Clonality: | Monoclonal |
| Clone number: | JE65-71 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| 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: 77 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within human CSDE1 aa 661-710/710. |
| Positive control: | K562 cell lysate, Jurkat cell lysate, Hela cell lysate, A431 cell lysate, Hela, rat bladder tissue, human lung tissue, human thyroid tissue, human colon carcinoma tissue, human skin tissue, human spleen tissue, human breast carcinoma tissue, human esophagus tissue, human placenta tissue, mouse liver tissue, MEF cell lysate, NIH/3T3 cell lysate, SiHa. |
| Subcellular location: | Cytoplasm, nucleolus. |
| Recommended Dilutions:
WB IF-Cell IHC-P |
1:500-1:5,000 1:200 1:200-1:400 |
| Uniprot #: | SwissProt: P19338 Human | P09405 Mouse | P13383 Rat |
| Alternative names: | C23 FLJ45706 MS1116 NCL Nucl NUCL_HUMAN Nucleolin Protein C23 |
Images
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Fig1:
Western blot analysis of Nucleolin on different lysates with Rabbit anti-Nucleolin antibody (HA721073) at 1/5,000 dilution. Lane 1: K562 cell lysate Lane 2: Jurkat cell lysate Lane 3: Hela cell lysate Lane 4: A431 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 77 kDa Observed band size: 100 kDa Exposure time: 2 minutes; 6% 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 (HA721073) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. |
|
Fig2:
Immunocytochemistry analysis of Hela cells labeling Nucleolin with Rabbit anti-Nucleolin antibody (HA721073) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Nucleolin antibody (HA721073) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (Alexa Fluor® 555) were used as the secondary antibody at 1/1,000 dilution. |
|
Fig3:
Immunohistochemical analysis of paraffin-embedded rat bladder tissue with Rabbit anti-Nucleolin antibody (HA721073) at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721073) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
|
Fig4:
Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-Nucleolin antibody (HA721073) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721073) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
|
Fig5:
Immunohistochemical analysis of paraffin-embedded human thyroid tissue with Rabbit anti-Nucleolin antibody (HA721073) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721073) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
|
Fig6:
Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-Nucleolin antibody (HA721073) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721073) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
|
Fig7:
Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Nucleolin antibody (HA721073) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721073) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
|
Fig8:
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Nucleolin antibody (HA721073) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721073) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
|
Fig9:
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Nucleolin antibody (HA721073) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721073) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
|
Fig10:
Immunohistochemical analysis of paraffin-embedded human esophagus tissue with Rabbit anti-Nucleolin antibody (HA721073) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721073) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
|
Fig11:
Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-Nucleolin antibody (HA721073) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721073) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
|
Fig12:
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Nucleolin antibody (HA721073) at 1/400 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721073) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
|
Fig13:
Western blot analysis of Nucleolin on different lysates with Rabbit anti-Nucleolin antibody (HA721073) at 1/500 dilution. Lane 1: MEF cell lysate Lane 2: NIH/3T3 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 77 kDa Observed band size: 100 kDa Exposure time: 30 seconds; 8% 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 (HA721073) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. |
|
Fig14:
Immunocytochemistry analysis of SiHa cells labeling Nucleolin with Rabbit anti-Nucleolin antibody (HA721073) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Nucleolin antibody (HA721073) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) were used as the secondary antibody at 1/1,000 dilution. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.