Anti-Semaphorin 3A antibody [EPR19367]

• IHC-P

Supplier Data

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.

• IHC-P

Supplier Data

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.

• IHC-P

Supplier Data

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.

• IHC-P

Supplier Data

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.

• IHC-P

Supplier Data

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.

• IHC-P

Supplier Data

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.

• IHC-P

Supplier Data

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.

• IP

Supplier Data

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.

All lanes:

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

Predicted band size: 89 kDa

false

• WB

Supplier Data

Western blot - Anti-Semaphorin 3A antibody [EPR19367] (AB199475)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Semaphorin 3A antibody [EPR19367] (ab199475) at 1/5000 dilution

Lane 1:

Human fetal brain lysate at 20 µg

Lane 2:

E14 rat embryo lysate at 20 µg

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

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Semaphorin 3A antibody [EPR19367] (AB199475)

Blocking/Dilution buffer : 5% NFDM/TBST.

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

All lanes:

Western blot - Anti-Semaphorin 3A antibody [EPR19367] (ab199475) at 1/1000 dilution

Lane 1:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 20 µg

Lane 2:

C6 (Rat glial tumor cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 89 kDa

Observed band size: 89 kDa

false

• WB

Supplier Data

Western blot - Anti-Semaphorin 3A antibody [EPR19367] (AB199475)

Blocking/Dilution buffer : 5% NFDM/TBST.

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

All lanes:

Western blot - Anti-Semaphorin 3A antibody [EPR19367] (ab199475) at 1/1000 dilution

Lane 1:

P0 mouse brain lysate at 20 µg

Lane 2:

Neuro-2a (Mouse neuroblastoma cell line) whole cell lysate at 20 µg

Lane 3:

Mouse kidney lysate at 20 µg

Lane 4:

Rat kidney lysate at 20 µg

Lane 5:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 89 kDa

Observed band size: 89 kDa

false