Recombinant Anti-GAP43 antibody [EP890Y] - Neuronal Marker (ab75810)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP890Y] to GAP43 - Neuronal Marker
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-P
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-GAP43 antibody [EP890Y] - Neuronal Marker
See all GAP43 primary antibodies -
Description
Rabbit monoclonal [EP890Y] to GAP43 - Neuronal Marker -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: SH-SY5Y, Human brain, Mouse brain, Rat brain, and PC-12 lysates. IHC-P: human cerebrum, mouse cerebrum, and rat cerebrum tissues. ICC/IF: Neuro-2a cells. Flow Cyt (intra): SH-SY5Y cells. IP: SH-SY5Y cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP890Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-GAP43 antibody [EP890Y] - Neuronal Marker (ab196324)
- HRP Anti-GAP43 antibody [EP890Y] - Neuronal Marker (ab196325)
- Alexa Fluor® 647 Anti-GAP43 antibody [EP890Y] - Neuronal Marker (ab196540)
- PE Anti-GAP43 antibody [EP890Y] - Neuronal Marker (ab208745)
- Anti-GAP43 antibody [EP890Y] - BSA and Azide free (ab219582)
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab75810 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
ICC/IF | (1) |
1/160.
For unpurified use at 1/500.
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WB | (2) |
1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 25 kDa).
For unpurified use at 1/100000 - 1/200000. The expression of GAP43 is undetectable in undifferentiated PC-12 cells in Western Blot (Ref: PMID: 2139463, PMID: 15969743) |
IP |
1/20 - 1/50.
|
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IHC-P | (5) |
1/3000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/500. |
Notes |
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Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/160. For unpurified use at 1/500.
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WB
1/1000. Detects a band of approximately 48 kDa (predicted molecular weight: 25 kDa). For unpurified use at 1/100000 - 1/200000. The expression of GAP43 is undetectable in undifferentiated PC-12 cells in Western Blot (Ref: PMID: 2139463, PMID: 15969743) |
IP
1/20 - 1/50. |
IHC-P
1/3000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/500. |
Target
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Function
This protein is associated with nerve growth. It is a major component of the motile "growth cones" that form the tips of elongating axons. -
Sequence similarities
Belongs to the neuromodulin family.
Contains 1 IQ domain. -
Post-translational
modificationsPhosphorylation of this protein by a protein kinase C is specifically correlated with certain forms of synaptic plasticity. -
Cellular localization
Cell membrane. Cell projection > growth cone membrane. Cell junction > synapse. Cytoplasmic surface of growth cone and synaptic plasma membranes. - Information by UniProt
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Database links
- Entrez Gene: 2596 Human
- Entrez Gene: 14432 Mouse
- Entrez Gene: 29423 Rat
- GenBank: NP_002036 Human
- Omim: 162060 Human
- SwissProt: P17677 Human
- SwissProt: P06837 Mouse
- SwissProt: P07936 Rat
see all -
Alternative names
- Axonal membrane protein GAP 43 antibody
- Axonal membrane protein GAP-43 antibody
- B 50 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAP43 antibody [EP890Y] - Neuronal Marker (ab75810)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebrum tissue sections labeling GAP43 with Purified ab75810 at 1:3000 dilution (0.07 µg/ml). Heat mediated antigen retrieval using Bond© Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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All lanes : Anti-GAP43 antibody [EP890Y] - Neuronal Marker (ab75810) (Purified)
Lane 1 : Human brain lysates
Lane 2 : Mouse brain lysates
Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 25 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted?This antibody fails to detect GAP43 in PC-12 cells which is positve as described in PMID: 21695168
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Immunocytochemistry/ Immunofluorescence analysis of Neuro-2a (Mouse neuroblastoma neuroblast) cells labeling GAP43 with Purified ab75810 at 1:160 dilution (1.4 µg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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ab75810 (Purified) at 1:20 dilution (1 µg) immunoprecipitating GAP43 in SH-SY5Y whole cell lysate.
Lane 1 (input): SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+): ab75810 & SH-SY5Y whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab75810 in SH-SY5Y whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAP43 antibody [EP890Y] - Neuronal Marker (ab75810)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebrum tissue sections labeling GAP43 with Purified ab75810 at 1:3000 dilution (0.07 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Intracellular Flow Cytometry analysis of SH-SY5Y (Human neuroblastoma epithelial cell) cells labeling GAP43 with Purified ab75810 at 1/20 dilution (10 µg/ml) (Red). Cells were fixed with 80% Methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAP43 antibody [EP890Y] - Neuronal Marker (ab75810)Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling GAP43 with Purified ab75810 at 1:3000 dilution (0.07 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Anti-GAP43 antibody [EP890Y] - Neuronal Marker (ab75810) at 1/50000 dilution (Purified) + SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 25 kDa
Observed band size: 48 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAP43 antibody [EP890Y] - Neuronal Marker (ab75810)This image is a courtesy of anonymous Abreview.
ab75810 (unpurified) staining GAP43 in Mouse ear tissue sections by Immunohistochemistry (Formalin/ PFA-fixed paraffin-embedded tissue sections). The sections were formaldehyde fixed, subjected to heat mediated antigen retrieval at pH 6 and blocked for 10 minutes at 25C. The primary antibody was diluted 1/500 and incubated with the sample for 1 hour at 25°C. An HRP polymer anti-rabbit IgG system was used undiluted, as the secondary antibody.
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Overlay histogram showing SH-SY5Y cells stained with ab75810 (unpurified) (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75810, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in SH-SY5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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ICC/IF image of ab75810 (unpurified) stained SKNSH cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab75810, 1/50 dilution) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG(H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (54)
ab75810 has been referenced in 54 publications.
- Feng S et al. Effects of combination treatment with transcranial magnetic stimulation and bone marrow mesenchymal stem cell transplantation or Raf inhibition on spinal cord injury in rats. Mol Med Rep 23:N/A (2021). PubMed: 33649786
- Zhu S et al. AAV2-mediated and hypoxia response element-directed expression of bFGF in neural stem cells showed therapeutic effects on spinal cord injury in rats. Cell Death Dis 12:274 (2021). PubMed: 33723238
- Ferdoushi A et al. Tumor innervation and clinical outcome in pancreatic cancer. Sci Rep 11:7390 (2021). PubMed: 33795769
- Nazir FH et al. Molecular forms of neurogranin in cerebrospinal fluid. J Neurochem 157:816-833 (2021). PubMed: 33249594
- Kim HN et al. A Western diet impairs CNS energy homeostasis and recovery after spinal cord injury: Link to astrocyte metabolism. Neurobiol Dis 141:104934 (2020). PubMed: 32376475