Anti-GAP43 antibody - Neuronal Marker (ab16053)

Rabbit polyclonal GAP43 antibody. Validated in WB, IHC, ICC/IF and tested in Mouse, Rat. Cited in 24 publication(s). Independently reviewed in 11 review(s).

Overview

  • Product name

    Anti-GAP43 antibody - Neuronal Marker
    See all GAP43 primary antibodies
  • Description

    Rabbit polyclonal to GAP43 - Neuronal Marker
  • Host species

    Rabbit
  • Specificity

    This antibody does not perform well in IHC-P (abreview 21090, 21089)
  • Tested applications

    Suitable for: ICC/IF, IHC (PFA fixed), WB, IHC-Frmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat
    Predicted to work with: Human, Chimpanzee
  • Immunogen

    Synthetic peptide corresponding to Rat GAP43 aa 200 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab16379)

  • Positive control

    • This antibody gave a positive signal in Rat dorsal root ganglion tissue that has been subjected to a spinal nerve ligation as well as Mouse spinal cord tissue lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab16053 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
ICC/IF 1/500.
IHC (PFA fixed) 1/300 - 1/1000.

PMID-28197545

WB Use a concentration of 1 µg/ml. Detects a band of approximately 43-53 kDa (predicted molecular weight: 43-53 kDa).Can be blocked with Rat GAP43 peptide (ab16379).
IHC-Fr 1/500.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • 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
      modifications

      Phosphorylation 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
    • Database links

    • Alternative names

      • 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
      • Nerve growth related peptide GAP43 antibody
      • NEUM_HUMAN antibody
      • Neural phosphoprotein B 50 antibody
      • Neural phosphoprotein B-50 antibody
      • Neuromodulin antibody
      • Neuron growth associated protein 43 antibody
      • PP46 antibody
      • Protein F1 antibody
      • QtrA-11580 antibody
      • QtrA-13071 antibody
      see all

    Images

    • ab16053 staining GAP43 in murine dorsal root ganglia by Immunocytochemistry/ Immunofluorescence.
      Cells were fixed with methanol, permeabilized using 0.1% saponin/PBS, blocked with 4% BSA for 30 minutes at 25°C and then incubated with ab16053 at a 1/500 dilution for 14 hours at 4°C. The secondary antibody used was a DyLight 488 conjugated goat anti-rabbit polyclonal used at a 1/100 dilution.

      See Abreview

    • All lanes : Anti-GAP43 antibody - Neuronal Marker (ab16053) at 1 µg/ml

      Lane 1 : spinal cord chung rat 1
      Lane 2 : spinal cord chung rat 2
      Lane 3 : spinal cord chung rat 3

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Dako Goat polyclonal to Rabbit IgG (HRP) at 1/2000 dilution

      Performed under reducing conditions.

      Predicted band size: 43-53 kDa
      Observed band size: 43 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 90 kDa (possible multimer)

    • Immunofluorescent staining obtained with ab16053 Rabbit polyclonal to GAP43 in rat dorsal root ganglion, 2 weeks after a spinal nerve injury. The staining is localised in the cytoplasm of some lesioned neurons (arrow in B) and in many axons (arrow heads in B). Image B (X40 objective) is a higher magnification of A (X20). The staining disappeared completely when the antibody was pre-incubated with the immunising (blocking) peptide (ab16379).

      Protocol details: Rats were intracardially perfused with 4% paraformaldehyde, DRG tissue was post-fixed overnight in the same fixative, cryoprotected in 20% sucrose and frozen in OCT. IHC free floating was performed on the fixed, frozen and cut (20µm sections) rat tissue. Primary antibody ab16053 was incubated overnight at toom temperature at 1/300. Secondary antibody Alexa fluor 488 was incubated at 1/1000 for 2 hours at room temperature.

    • Anti-GAP43 antibody - Neuronal Marker (ab16053) at 1 µg/ml + Spinal Cord (Mouse) Tissue Lysate at 10 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 43-53 kDa
      Observed band size: 43 kDa why is the actual band size different from the predicted?
      Additional bands at: 100 kDa, 50 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 8 minutes

    References

    This product has been referenced in:

    • van der Merwe Y  et al. Matrix-bound nanovesicles prevent ischemia-induced retinal ganglion cell axon degeneration and death and preserve visual function. Sci Rep 9:3482 (2019). Read more (PubMed: 30837658) »
    • Li L  et al. Ascorbic Acid Facilitates Neural Regeneration After Sciatic Nerve Crush Injury. Front Cell Neurosci 13:108 (2019). Read more (PubMed: 30949031) »
    See all 29 Publications for this product

    Customer reviews and Q&As

    1-10 of 15 Abreviews or Q&A

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (spinal cord)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris)
    Loading amount
    10 µg
    Specification
    spinal cord
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 20°C

    Abcam user community

    Verified customer

    Submitted May 17 2019

    Answer

    We would recommend using ab16053 in place of the discontinued item. Ab16053 has been tested in and is guaranteed in IF on mouse samples. This product has also been featured in 5 publications which we are aware of and has excellent Abreviews. More information about this may be found at: https://www.abcam.com/gap43-antibody-neuronal-marker-ab16053.html Abcam also carries a number of reagents for IF staining, including serums and blocking reagents, mounting media and background reducing buffers. Our BrightMount Plus Aqueous Mounting Medium for Fluorescent Staining (ab103748; https://www.abcam.com/BrightMount-Plus-25ml-Aqueous-Mounting-Medium-Anti-fading-for-Fluorescent-Staining-ab103748.html) is specifically designed for use in IF and would protect against photo-bleaching of your samples.

    Read More
    This product is known to not work in this application or species.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (mouse brain (hippocampus))
    Specification
    mouse brain (hippocampus)
    Fixative
    4% PFA fixed
    Antigen retrieval step
    None
    Permeabilization
    Yes - 0,1% Saponin
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Nov 19 2012

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (mouse lung (bronchus))
    Specification
    mouse lung (bronchus)
    Fixative
    Methanol
    Permeabilization
    Yes - 0,1% Saponin
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Nov 12 2012

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (brain tissue)
    Loading amount
    10 µg
    Specification
    brain tissue
    Gel Running Conditions
    Reduced Non-Denaturing (Native) (5-16% gel)
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Nov 09 2012

    Answer

    Thank you for your reply.

    Your protocol looks good. The one recommendation I could make would be to increase the dilution of the primary and secondary (especially) to see if that improves the staining.
    You can try the primary at 1:250 and the secondary at 1:200 and see if that helps.

    If this does not improve the staining, we would be happy to replace or refund the product. Please let me know if this does not help and what you would prefer as compensation.

    Read More

    Answer

    Thank you for calling us and for alerting us to the problem you are experiencing with anti-GAP43 antibody (ab16053). We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

    As we discussed on the phone, Ihave attachedour questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary.

    If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

    I look forward to receiving your reply.

    Read More
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (dorsal root ganglia)
    Specification
    dorsal root ganglia
    Fixative
    Methanol
    Permeabilization
    Yes - 0,1% Saponin / PBS
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Dec 12 2011

    This product is known to not work in this application or species.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (in situ spinal cord)
    Specification
    in situ spinal cord
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid pH6
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

    Mr. Carl Hobbs

    Verified customer

    Submitted Nov 02 2010

    This product is known to not work in this application or species.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Head)
    Specification
    Head
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid pH6
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

    Mr. Carl Hobbs

    Verified customer

    Submitted Nov 02 2010

    1-10 of 15 Abreviews or Q&A

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