Recombinant
RabMAb

Anti-Semaphorin 3A antibody [EPR19367] (ab199475)

Overview

  • Product name
    Anti-Semaphorin 3A antibody [EPR19367]
    See all Semaphorin 3A primary antibodies
  • Description
    Rabbit monoclonal [EPR19367] to Semaphorin 3A
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse Semaphorin 3A aa 500-750. The exact sequence is proprietary.
    Database link: O08665

  • Positive control
    • WB: Human fetal brain lysate; E14 rat embryo lysate; P0 mouse brain lysate; mouse and rat kidney lysates; Neuro-2a, NIH/3T3, PC-12, and C6 whole cell lysates. IHC-P: Human kidney, colon cancer and vascular smooth muscle of human skin tissues; Mouse kidney, vascular smooth muscle of mouse skin, and cerebral cortex of mouse E14 tissues; Rat heart tissue. IP: PC-12 whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab199475 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
WB 1/1000. Detects a band of approximately 89 kDa (predicted molecular weight: 89 kDa).
IHC-P 1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/40.

Target

  • Relevance
    One family of inhibitory axon guidance molecules is the semaphorins. The semaphorins include secreted, transmembrane, and GPI anchored extracellular molecules that are involved in regulating axon guidance by inhibiting axons from growing toward incorrect targets. Semaphorin 3A (Sema3A) may play a particularly interesting role in limiting axon regeneration since it is expressed in meningeal fibroblasts that invade the injured spinal cord and surround the glial scar. In addition, the Sema3A co receptors, Neuropilin 1 and Plexin A1, are expressed on axons that regenerate up to the injured region, but do not cross this Sema3A containing region. Thus, Sema3A and its co receptors may have important roles in regulating axon guidance during neuronal development and after neuronal injury.
  • Cellular localization
    Secreted
  • Database links
  • Alternative names
    • Coll 1 antibody
    • Coll1 antibody
    • Collapsin 1 antibody
    • HH16 antibody
    • Hsema I antibody
    • Hsema III antibody
    • Sema domain immunoglobulin domain (Ig) short basic domain secreted (semaphorin) 3A antibody
    • sema domain, immunoglobulin domain (Ig), short basic domain, secreted, (semaphorin) 3A antibody
    • Sema III antibody
    • SEMA1 antibody
    • SEMA1-PEN antibody
    • SEMA3A antibody
    • SEMAD antibody
    • SEMAIII antibody
    • SEMAL antibody
    • Semaphorin 3A precursor antibody
    • Semaphorin III antibody
    • semaphorin L antibody
    • Semaphorin like growth cone guidance homolog of chick collapsin antibody
    • Semaphorin-D antibody
    • Semaphorin3A antibody
    • SemD antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Semaphorin 3A with ab199475 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on vascular smooth muscle and podocytes of human kidney [PMID: 25475434]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

  • All lanes : Anti-Semaphorin 3A antibody [EPR19367] (ab199475) at 1/5000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : E14 rat embryo lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 89 kDa
    Observed band size: 89 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Semaphorin 3A antibody [EPR19367] (ab199475) at 1/1000 dilution

    Lane 1 : P0 mouse brain lysate
    Lane 2 : Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Rat kidney lysate
    Lane 5 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 89 kDa
    Observed band size: 89 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1, 2, 3 and 4: 15 seconds; Lane 5: 3 seconds.

  • All lanes : Anti-Semaphorin 3A antibody [EPR19367] (ab199475) at 1/1000 dilution

    Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 2 : C6 (Rat glial tumor cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 89 kDa
    Observed band size: 89 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1:10 seconds; Lane 2:3 minutes.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling Semaphorin 3A with ab199475 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on vascular smooth muscle of human colon cancer [PMID: 18483621]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Semaphorin 3A with ab199475 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on vascular smooth muscle and podocytes of mouse kidney was observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

  • Immunohistochemical analysis of paraffin-embedded cerebral cortex of mouse E14 tissue labeling Semaphorin 3A with ab199475 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on perineural small blood vessels of mouse E14 cerebral cortex [PMID: 12879061]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

  • Immunohistochemical analysis of paraffin-embedded rat heart tissue labeling Semaphorin 3A with ab199475 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining on vascular smooth muscle of rat heart. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

  • Immunohistochemical analysis of paraffin-embedded human skin tissue labeling Semaphorin 3A with ab199475 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Weak cytoplasmic staining on vascular smooth muscle of human skin; epidermis is negative [PMID: 19443185]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

  • Immunohistochemical analysis of paraffin-embedded mouse skin tissue labeling Semaphorin 3A with ab199475 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Weak cytoplasm staining on vascular smooth muscle of mouse skin was observed, staining on the epidermis is negative [PMID: 19443185]. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

  • Semaphorin 3A was immunoprecipitated from 0.35 mg of PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate with ab199475 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab199475 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.

    Lane 1: PC-12 whole cell lysate, 10 μg (Input).

    Lane 2: ab199475 IP in PC-12 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab199475 in PC-12 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

    Staining pattern is consistent with what has been described in the literature, PMID: 23469280.

References

ab199475 has not yet been referenced specifically in any publications.

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

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