Human/Mouse/Rat Semaphorin 5A Antibody

Name: Human/Mouse/Rat Semaphorin 5A Antibody
Brand: R&D Systems
SKU: AF5896
Price: 459 USD

Catalog #: AF5896
2 Citations Datasheet / COA / SDS

Discontinued Product

AF5896 has been discontinued.

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Detection of Mouse Semaphorin 5A by Western Blot.

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Product Details

Citations (2)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse/Rat Semaphorin 5A Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human, mouse, and rat Semaphorin 5A in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human (rh) Sema3B, rhSema4C, and rhSema6A is observed.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human Semaphorin 5A
Glu23-Thr765
Accession # AAC09473

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

Immunocytochemistry

5-15 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Mouse Semaphorin 5A by Western Blot. Western blot shows lysates of beta TC-6 mouse beta cell insulinoma cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat Semaphorin 5A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5896) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Semaphorin 5A at approximately 135 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

Immunocytochemistry

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Semaphorin 5A in beta TC‑6 Mouse Cell Line. Semaphorin 5A was detected in immersion fixed beta TC-6 mouse beta cell insulinoma cell line using Sheep Anti-Human/Mouse/Rat Semaphorin 5A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5896) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry

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Semaphorin 5A in Human Pancreas. Semaphorin 5A was detected in immersion fixed paraffin-embedded sections of human pancreas using Sheep Anti-Human/Mouse/Rat Semaphorin 5A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5896) at 5 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hemotoxylin (blue). Specific staining was localized to the plasma membranes of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Semaphorin 5A

Semaphorin 5A (Sema5A, previously called SemaF) is a 140 kDa protein of the semaphorin family of axon guidance molecules (1‑4). Class 5 semaphorins are type I transmembrane glycoproteins with an N-terminal Sema domain and multiple juxtamembrane type 1 thrombospondin (TSP) repeats within their extracellular domains (1‑3). Sema5A is expressed developmentally in oligodendrocytes, neuroepithelial cells surrounding retinal axons, the base of limb buds, the cardiac atrial septum and endocardial cushions, and the mesoderm surrounding cranial vessels (4‑6). The human Sema5A cDNA encodes a 22 amino acid (aa) signal sequence, a 946 aa extracellular domain (ECD), a 22 aa transmembrane sequence and an 85 aa cytoplasmic portion. Within aa 23‑765, which includes the sema domain and four of the seven TSP repeats, human Sema5A shares 93% aa identity with corresponding mouse, rat, and canine sequences. Semaphorins typically transduce signals through transmembrane plexins (1, 2). The sema domain of Sema5A binds plexin B3, triggering signaling via HGF R/c-Met (7). Both Sema5A and plexin B3 are expressed postnatally during differentiation and migration of central nervous system oligodendrocytes. However, plexin B3 is not significantly expressed prenatally and therefore unlikely to be the Sema5A receptor during development (7, 8). The Sema5A TSP repeats interact with either heparin sulfate or chondroitin sulfate proteoglycans (HSPG, CSPG) (9). HSPG interaction promotes attraction, while CSPG interaction promotes repulsion and is essential for axon fasciculation, independent of plexin B3 (9, 10). Sema5A mutations have been implicated in the genetic syndrome, cri-du-chat, while some polymorphisms may increase risk for neurodegenerative diseases such as Parkinson’s (3, 11). Sema5A expression may be upregulated in metastatic cancer cells and downregulated in autism (12, 13).

References

Flannery, E. and M. Duman-Scheel (2009) Curr. Drug Targets 10:611.
Zhou, Y. et al. (2008) Trends Biol. Sci. 33:161.
Simmons, A.D. et al. (1998) Biochem. Biophys. Res. Commun. 242:685.
Oster, S.F. et al. (2003) Development 130:775.
Goldberg, J.L. et al. (2004) J. Neurosci. 24:4989.
Fiore, R. et al. (2005) Mol. Cell. Biol. 25:2310.
Artigiani, S. et al. (2004) EMBO Rep. 5:710.
Worzfeld, T. et al. (2004) Eur. J. Neurosci. 19:2622.
Kantor, D.B. et al. (2004) Neuron 44:961.
Hilario, J.D. et al. (2008) Dev. Biol. 326:190.
Lin, L. et al. (2009) Trends Neurosci. 32:142.
Pan, G-Q. et al. (2009) World J. Gastroenterol. 15:2800.
Melin, M. et al. (2006) Neuropsychobiology 54:64.

Entrez Gene IDs

9037 (Human); 20356 (Mouse); 310207 (Rat)

Alternate Names

FLJ12815; sema domain, seven thrombospondin repeats (type 1 and type 1-like); Sema F; SEMA5A; SEMAF; Semaphorin 5A; semaphorin F; semaphorin-5A; Semaphorin-F; semF; transmembrane domain (TM) and short cytoplasmic domain, (semaphorin) 5A

Product Datasheets

Product Datasheet

COA

SDS

Related Research Areas

Citations for Human/Mouse/Rat Semaphorin 5A Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

2 Citations: Showing 1 - 2
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Plexin-B3 expression stimulates MET signaling, breast cancer stem cell specification, and lung metastasis.
Authors: Zuo Q, Yang Y, Lyu Y, Yang C, Chen C, Salman S, Huang T, Wicks E, Jackson W, Datan E, Qin W, Semenza G
Cell Rep, 2023-02-28;42(3):112164.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
A mutant p53-dependent embryonic stem cell gene signature is associated with augmented tumorigenesis of stem cells
Authors: G Koifman, Y Shetzer, S Eizenberge, H Solomon, R Rotkopf, A Molchadsky, G Lonetto, N Goldfinger, V Rotter
Cancer Res., 2018;0(0):.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC