GAP43 Recombinant Rabbit Monoclonal Antibody [SC60-06]
cat.: ET1610-94
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, IF-Tissue, IHC-P, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | SC60-06 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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: 25 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within Human GAP43 aa 189-238 / 238. |
| Positive control: | SH-SY5Y cell lysate, Neuro-2a cell lysate, mouse brain tissue lysate, rat brain tissue lysate, mouse brain tissue, Neuro-2a, rat hippocampus tissue lysate, mouse hippocampus tissue lysate, SH-SY5Y, human glioma tissue. |
| Subcellular location: | Cell membrane, growth cone membrane, filopodium membrane, Cytoplasm, synapse, perikaryon, dendrite, axon. |
| Recommended Dilutions:
WB IF-Cell IF-Tissue IHC-P FC IP |
1:5,000-1:100,000 1:500 1:100-1:500 1:5,000-1:15,000 1:1,000 Use at an assay dependent concentration. |
| Uniprot #: | SwissProt: P17677 Human | P06837 Mouse | P07936 Rat |
| Alternative names: | Axonal membrane protein GAP 43 Axonal membrane protein GAP-43 B 50 Calmodulin binding protein P 57 F1 GAP 43 GAP43 Growth Associated Protein 43 Growth-associated protein 43 Nerve Growth Related Peptide Nerve growth related peptide GAP43 NEUM_HUMAN Neural phosphoprotein B 50 Neural phosphoprotein B-50 Neuromodulin Neuron growth associated protein 43 PP46 Protein F1 QtrA-11580 QtrA-13071 |
Images
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Fig1:
Western blot analysis of GAP43 on different lysates with Rabbit anti-GAP43 antibody (ET1610-94) at 1/100,000 dilution and competitor's antibody at 1/100,000 dilution. Lane 1: SH-SY5Y cell lysate Lane 2: A549 cell lysate (negative) Lane 3: Neuro-2a cell lysate Lane 4: Mouse lung tissue lysate (negative) Lane 5: Mouse brain tissue lysate Lane 6: Rat brain tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 25 kDa Observed band size: 43/45 kDa Exposure time: 3 minutes 20 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 (ET1610-94) at 1/100,000 dilution and competitor's antibody at 1/100,000 dilution were 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:
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-GAP43 antibody (ET1610-94) at 1/15,000 dilution and competitor's antibody at 1/3,000 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 (ET1610-94) at 1/15,000 dilution and competitor's antibody at 1/3,000 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. |
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Fig3:
Immunocytochemistry analysis of Neuro-2a cells labeling GAP43 with Rabbit anti-GAP43 antibody (ET1610-94) at 1/500 dilution. Cells were fixed in 100% precooled methanol 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-GAP43 antibody (ET1610-94) at 1/500 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:
Western blot analysis of GAP43 on different lysates with Rabbit anti-GAP43 antibody (ET1610-94) at 1/5,000 dilution. Lane 1: SH-SY5Y cell lysate Lane 2: Neuro-2a cell lysate Lane 3: Mouse brain tissue lysate Lane 4: Rat brain tissue lysate Lane 5: Rat hippocampus tissue lysate Lane 6: Mouse hippocampus tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 25 kDa Observed band size: 43/45 kDa Exposure time: 24 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 (ET1610-94) at 1/5,000 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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Fig5:
Western blot analysis of GAP43 on different lysates with Rabbit anti-GAP43 antibody (ET1610-94) at 1/20,000 dilution. Lane 1: SH-SY5Y-si NT cell lysate Lane 2: SH-SY5Y-si GAP43 cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 25 kDa Observed band size: 45 kDa Exposure time: 30 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 (ET1610-94) at 1/20,000 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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Fig6:
Intracellular Flow Cytometry analysis of SH-SY5Y labeling GAP43 with purified ET1610-94 at 1/1,000 dilution (1 µg/ml) (red). Cells were fixed with 4% PFA and permeabilised with 90% methanol. Rabbit monoclonal IgG (green) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (black) were used as the unlabeled control. A Goat anti-rabbit IgG iFluor™ 488 (HA1121)(1/1,000 dilution) was used as the secondary antibody. |
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Fig7:
Immunohistochemical analysis of paraffin-embedded human glioma tissue with Rabbit anti-GAP43 antibody (ET1610-94) at 1/15,000 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 (ET1610-94) at 1/15,000 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. |
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Fig8:
Flow cytometric analysis of Neuro-2a cells labeling GAP43. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1610-94, 1μg/mL) (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). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.