Name: Anti-Semaphorin 3A antibody [EPR19367]
Brand: RabMAb
SKU: ab199475
Rating: 4 (2 reviews)

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Western blot - 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

Carrier Free

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.

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, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19367
Isotype

IgG
Research areas

Alternative Versions
- Anti-Semaphorin 3A antibody [EPR19367] - Low endotoxin, Azide free (ab222497)
- Anti-Semaphorin 3A antibody [EPR19367] - BSA and Azide free (ab271964)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

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.

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.
Target information above from: UniProt accession Q14563 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 10371 Human
- Entrez Gene: 20346 Mouse
- Entrez Gene: 29751 Rat
- Omim: 603961 Human
- SwissProt: Q14563 Human
- SwissProt: O08665 Mouse
- SwissProt: Q63548 Rat
- Unigene: 252451 Human
- Unigene: 372039 Mouse
- Unigene: 483774 Mouse
- Unigene: 44410 Rat
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

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)

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)

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)

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)

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)

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)

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)

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)

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)

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)

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)

Western blot protocols
Immunoprecipitation protocols
Immunohistochemistry protocols

Click here to view the general protocols

Datasheet download

Download

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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1-2 of 2 Abreviews or Q&A

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (nhek)

Permeabilization

Yes - triton 0.5%

Specification

nhek

Blocking step

BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Mar 03 2022

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

Katrin Mooslehner

Verified customer

Submitted Mar 05 2021

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Domain

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (1)

Customer reviews and Q&As

Immunocytochemistry/ Immunofluorescence abreview for Anti-Semaphorin 3A antibody [EPR19367]

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