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GAP43 Recombinant Rabbit Monoclonal Antibody [SC60-06]

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Specification

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Catalog# ET1610-94

GAP43 Recombinant Rabbit Monoclonal Antibody [SC60-06]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • IP

  • FC(Intra)

Reactivity

  • Human

  • Mouse

  • Rat

Predicted reactivity

  • Cynomolgus monkey

  • Pig

Predicted species support after-sales service
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

GAP43 Recombinant Rabbit Monoclonal Antibody [SC60-06]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human GAP43 aa 189-238 / 238.

Species Reactivity

Human, Mouse, Rat (Predicted: Cynomolgus monkey, Pig)

Predicted species support after-sales service

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, IP, FC(Intra)

Molecular Weight

Predicted band size: 25 kDa

Positive Control

SH-SY5Y cell lysate, Neuro-2a cell lysate, mouse brain tissue lysate, rat brain tissue lysate, human glioma tissue, Neuro-2a, rat hippocampus tissue lysate, mouse hippocampus tissue lysate, mouse brain tissue, rat brain tissue, rat hippocampus tissue, SH-SY5Y.

Conjugation

unconjugated

Clone Number

SC60-06

RRID

AB_3069981

Product Features

Form

Liquid

Concentration

Storage Instructions

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.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:20,000-1:100,000

  • IF-Cell

  • 1:500

  • IF-Tissue

  • 1:500-1:1,000

  • IHC-P

  • 1:10,000-1:15,000

  • FC(Intra)

  • 1:1,000

  • IP

  • Use at an assay dependent concentration.

Target

Function

GAP43, is a nervous tissue-specific cytoplasmic protein that can be attached to the membrane via a dual palmitoylation sequence on cysteines 3 and 4. This sequence targets GAP43 to lipid rafts. It is a major protein kinase C (PKC) substrate and is considered to play a key role in neurite formation, regeneration, and plasticity. The role of GAP-43 in CNS development is not limited to effects on axons: It is also a component of the centrosome, and differentiating neurons that do not express GAP-43 show mislocalization of the centrosome and mitotic spindles, particularly in neurogenic cell divisions. GAP43, the consensus choice for its designation, is a nervous system-specific protein that is attached to the membrane via a dual palmitoylation sequence on cysteines 3 and 4, though it can exist in the non-bound form in the cytoplasm. This dual sequence enables the association of phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2] or PIP2, with actin, facilitating the latter's polymerization thereby regulating neuronal structure. GAP-43 is also a protein kinase C (PKC) substrate. Phosphorylation of serine-41 on GAP-43 by PKC regulates neurite formation, regeneration, and synaptic plasticity.

Background References

1. D\'Agostino G et al. Prolyl endopeptidase-deficient mice have reduced synaptic spine density in the CA1 region of the hippocampus, impaired LTP, and spatial learning and memory. Cereb Cortex 23:2007-14 (2013).

2. Figueroa JD et al. Metabolomics uncovers dietary omega-3 fatty acid-derived metabolites implicated in anti-nociceptive responses after experimental spinal cord injury. Neuroscience 255C:1-18 (2013).

Sequence Similarity

Belongs to the neuromodulin family.

Post-translational Modification

Phosphorylated (By similarity). Phosphorylation of this protein by a protein kinase C is specifically correlated with certain forms of synaptic plasticity (By similarity).; Palmitoylated by ZDHHC3 (By similarity). Palmitoylation is regulated by ARF6 and is essential for plasma membrane association and axonal and dendritic filopodia induction. Deacylated by LYPLA2.

Subcellular Location

Cell membrane, growth cone membrane, filopodium membrane, Cytoplasm, synapse, perikaryon, dendrite, axon.

UNIPROT

SwissProt: P17677 Human

SwissProt: P06837 Mouse

SwissProt: P07936 Rat

Synonyms

Axonal membrane protein GAP 43 antibody

Axonal membrane protein GAP-43 antibody

B 50 antibody

Calmodulin binding protein P 57 antibody

F1 antibody

GAP 43 antibody

GAP43 antibody

Growth Associated Protein 43 antibody

Growth-associated protein 43 antibody

Nerve Growth Related Peptide antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of GAP43 on different lysates with Rabbit anti-GAP43 antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) at 1/100,000 dilution and competitor's antibody at 1/100,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate<br />Lane 2: A549 cell lysate (negative)<br />Lane 3: Neuro-2a cell lysate<br />Lane 4: Mouse lung tissue lysate (negative)<br />Lane 5: Mouse brain tissue lysate<br />Lane 6: Rat brain tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 25 kDa<br />Observed band size: 43/45 kDa<br /><br />Exposure time: 3 minutes 20 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) 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 (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    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; 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 (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.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Glioma<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>, 1/10,000; competitor's antibody, 1/3,000, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Glioma
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: ET1610-94, 1/10,000; competitor's antibody, 1/3,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Immunocytochemistry analysis of Neuro-2a cells labeling GAP43 with Rabbit anti-GAP43 antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) at 1/500 dilution.<br /><br />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 (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    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 (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.

  • Western blot analysis of GAP43 on different lysates with Rabbit anti-GAP43 antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) at 1/20,000 dilution.<br /><br />Lane 1: SH-SY5Y cell lysate<br />Lane 2: Neuro-2a cell lysate<br />Lane 3: Mouse brain tissue lysate<br />Lane 4: Rat brain tissue lysate<br />Lane 5: Rat hippocampus tissue lysate<br />Lane 6: Mouse hippocampus tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 25 kDa<br />Observed band size: 43/45 kDa<br /><br />Exposure time: 24 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of GAP43 on different lysates with Rabbit anti-GAP43 antibody (ET1610-94) at 1/20,000 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 1/20,000 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of GAP43 on different lysates with Rabbit anti-GAP43 antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) at 1/20,000 dilution.<br /><br />Lane 1: SH-SY5Y-si NT cell lysate<br />Lane 2: SH-SY5Y-si GAP43 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 25 kDa<br />Observed band size: 45 kDa<br /><br />Exposure time: 30 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    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.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Mouse<br />Tissue: Hippocampus<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>, 1/10,000, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Hippocampus
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: ET1610-94, 1/10,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Rat<br />Tissue: Hippocampus<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>, 1/15,000, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Rat
    Tissue: Hippocampus
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: ET1610-94, 1/15,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Immunofluorescence analysis of paraffin-embedded mouse brain tissue labeling GAP43 with Rabbit anti-GAP43 antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>) at 1/200 dilution.<br /><br />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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

    Immunofluorescence analysis of paraffin-embedded mouse brain tissue labeling GAP43 with Rabbit anti-GAP43 antibody (ET1610-94) at 1/200 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1610-94, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

  • Intracellular Flow Cytometry analysis of SH-SY5Y labeling GAP43 with purified <a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a> at 1/1,000 dilution (1 µg/ml) (red).<br /><br />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&trade; 488 (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>)(1/1,000 dilution) was used as the secondary antibody.

    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.

  • Flow cytometric analysis of Neuro-2a cells labeling GAP43.<br /><br />Cells were fixed with 4% PFA and permeabilised with 90% methanol. Then stained with the primary antibody (<a href="/products/ET1610-94" style="font-weight: bold;text-decoration: underline;">ET1610-94</a>, 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&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Neuro-2a cells labeling GAP43.

    Cells were fixed with 4% PFA and permeabilised with 90% methanol. 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).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

Products with the same target and pathway

metafield image

GAP43 Rabbit Polyclonal Antibody

Application: WB,IF-Cell,IHC-P,FC

Reactivity: Human,Mouse,Rat

Conjugate: unconjugated