WB
Western blot analysis of Nestin on mouse brain (P0) tissue lysates with Rat anti-Nestin antibody at 1/2,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 6 seconds; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rat IgG H&L - HRP Secondary Antibody at 1/5,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rat anti-Nestin antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 at 1/1,000 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.IHC
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rat anti-Nestin antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 at 1/1,000 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.IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rat anti-Nestin antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 at 1/1,000 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.ICC/IF
Immunocytochemistry analysis of C2C12 cells labeling Nestin with Rat anti-Nestin antibody at 1/1,000 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 Rat anti-Nestin antibody at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rat IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.IF-F
Immunofluorescence analysis of frozen E14.5 mouse embryo tissue with Rat anti-Nestin antibody at 1/1,000 dilution. The section was not undergone antigen retrieval. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (green) at 1/1,000 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rat IgG H&L (488) was used as the secondary antibody at 1/500 dilution. Nuclei were counterstained with DAPI (blue).| Product Name | Nestin Recombinant Antibody - Rat IgG1 (Chimeric) |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within mouse Nestin aa 700-1,000. |
| Clonality | Monoclonal |
|---|---|
| Tested Applications | ICC/IFIF-FIHCWB |
| WB:1:2000 IHC:1:1000 ICC/IF:1:1000-1:10000 IF-F:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | Nes |
|---|---|
| Gene Synonyms | RC2 Marc2 ESTM46 Ifaprc2 |
| Gene Full Name | nestin |
| Gene Summary | Predicted to enable CCR5 chemokine receptor binding activity and intermediate filament binding activity. Acts upstream of or within cell projection morphogenesis; negative regulation of neuron apoptotic process; and nervous system development. Located in cytoplasm and intermediate filament. Is expressed in several structures, including central nervous system; embryo mesenchyme; eye; foregut-midgut junction; and metanephros. Orthologous to human NES (nestin). [provided by Alliance of Genome Resources, Jul 2025] |
| Alternative Names | ESTM 46 antibody FLJ 21841 antibody FLJ21841 antibody Intermediate filament protein antibody Nbla00170 antibody nes antibody NEST_HUMAN antibody Nestin antibody |
| Molecular Weight(MW) | 207kDa(Observed band size: 300kDa) |
| Cellular Localization | Cytoplasm, Intermediate filament. |
WB
Western blot analysis of Nestin on mouse brain (P0) tissue lysates with Rat anti-Nestin antibody at 1/2,000 dilution. Lysates/proteins at 20 µg/Lane. Exposure time: 6 seconds; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rat IgG H&L - HRP Secondary Antibody at 1/5,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rat anti-Nestin antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 at 1/1,000 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.
IHC
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rat anti-Nestin antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 at 1/1,000 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rat anti-Nestin antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 at 1/1,000 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.
ICC/IF
Immunocytochemistry analysis of C2C12 cells labeling Nestin with Rat anti-Nestin antibody at 1/1,000 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 Rat anti-Nestin antibody at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rat IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
IF-F
Immunofluorescence analysis of frozen E14.5 mouse embryo tissue with Rat anti-Nestin antibody at 1/1,000 dilution. The section was not undergone antigen retrieval. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (green) at 1/1,000 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rat IgG H&L (488) was used as the secondary antibody at 1/500 dilution. Nuclei were counterstained with DAPI (blue).| Application Notes | WB:1:2000 IHC:1:1000 ICC/IF:1:1000-1:10000 IF-F:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
