Antibodies, Proteins, Kits and Reagents for Life Science

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 & Species

Application	Species
IHC-P	Mouse Rat Human
IP	Rat
WB	Mouse Rat Human

See all applications and species data

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

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)