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RecombinantRabMAb

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

Reviews (2) Submit a question References (1)
Western blot - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
  • Western blot - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
  • Western blot - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
  • Immunoprecipitation - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
  • Anti-Semaphorin 3A antibody [EPR19367] (ab199475)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19367] to Semaphorin 3A
  • Suitable for: WB, IHC-P, IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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. This information is proprietary to Abcam and/or its suppliers.

  • 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

    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
    For more information see here.

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

Properties

Applications

The Abpromise guarantee

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)
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.
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

  • Function

    Involved in the development of the olfactory system and in neuronal control of puberty. Induces the collapse and paralysis of neuronal growth cones. Could serve as a ligand that guides specific growth cones by a motility-inhibiting mechanism. Binds to the complex neuropilin-1/plexin-1.

  • Involvement in disease

    Hypogonadotropic hypogonadism 16 with or without anosmia

  • Sequence similarities

    Belongs to the semaphorin family.
    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 PSI domain.
    Contains 1 Sema domain.

  • Domain

    Strong binding to neuropilin is mediated by the carboxy third of the protein.

  • Cellular localization

    Secreted.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • Coll 1 antibody
    • Coll1 antibody
    • Collapsin 1 antibody
    • HH16 antibody
    • Hsema I antibody
    • Hsema III antibody
    • SEM3A_HUMAN 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-3A antibody
    • Semaphorin-D antibody
    • Semaphorin3A antibody
    • SemD antibody
    see all

Images

  • Western blot - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    Western blot - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)

    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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    Western blot - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    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.

  • Western blot - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    Western blot - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)

    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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)

    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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)

    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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)

    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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)

    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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Semaphorin 3A antibody [EPR19367] (ab199475)

    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.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    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 Detection Reagent (HRP) (ab131366), was used for detection 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.

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

References (1)

Publishing research using ab199475? Please let us know so that we can cite the reference in this datasheet.

ab199475 has been referenced in 1 publication.

  • Wu JH  et al. Semaphorin-3A protects against neointimal hyperplasia after vascular injury. EBioMedicine 39:95-108 (2019). ICC/IF ; Human . PubMed: 30579864

Customer reviews and Q&As

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Western blot abreview for Anti-Semaphorin 3A antibody [EPR19367]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Xenopus laevis Tissue lysate - whole (brain)
Gel Running Conditions
Reduced Denaturing (4-12% BisTris)
Loading amount
15 µg
Treatment
125uM and 250uM Sema3A morpholino
Specification
brain
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Katrin Mooslehner

Verified customer

Submitted Mar 05 2021

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