Western Blotting (WB), Flow Cytometry (FACS), Immunocytochemistry (ICC)
Specificity
The antibody Hs-14 reacts with VCP (valosin-containing protein) a 220 kDa intra-acrosomal protein.
Cross-Reactivity (Details)
Mouse, Human
Purification
Purified by sequential steps of physicochemical fractionation (differential precipitation and solid-phase chromatography methods).
Purity
> 95 % (by SDS-PAGE)
Immunogen
Freshly ejaculated human sperms were washed in PBS and extracted in 3% acetic acid, 10% glycerol, 30 mM benzaminidine. The acid extract was dialyzed against 0.2% acetic acid and subsequently used for immunization.
Immunocytochemistry: Recommended dilution: 10 μg/mL, membrane permeabilization (acetone) is essential. The antibody Hs-14 is designed for quantitative immunofluorescence analysis of pathological sperms (excellent tool for laboratories of assisted reproduction when optimal method of fertilization is sought). Flow cytometry: Intraacrosomal staining, recommended dilution: 3-12 μg/mL.
Restrictions
For Research Use only
Concentration
1 mg/mL
Buffer
Tris buffered saline (TBS), pH 8.0, 15 mM sodium azide
Preservative
Sodium azide
Precaution of Use
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice
Do not freeze.
Storage
4 °C
Storage Comment
Store at 2-8°C. Do not freeze.
Peknicova, Chladek, Hozak: "Monoclonal antibodies against sperm intra-acrosomal antigens as markers for male infertility diagnostics and estimation of spermatogenesis." in: American journal of reproductive immunology (New York, N.Y. : 1989), Vol. 53, Issue 1, pp. 42-9, (2005) (PubMed).
Chládek, P?knicová, Capková, Geussová, Teplá, Madar: "[Use of human sperm protein monoclonal antibodies in the diagnosis of sperm pathology and selection of a suitable assisted reproduction method for fertilization]." in: Ceská gynekologie / Ceská lékarská spolecnost J. Ev. Purkyne, Vol. 65, Issue 1, pp. 28-32, (2000) (PubMed).
Valosin containing protein,VCP (valosin-containing protein), also known as p97, TERA, ALS14, IBMPFD, HEL-220, IBMPFD1, or HEL-S-70, is a member of a protein family that includes putative ATP-binding proteins involved in vesicle transport and fusion, 26S proteasome function, and assembly of peroxisomes. VCP is a structural protein that associates with clathrin and heat-shock protein Hsc70, to form a complex. It has been implicated in a number of cellular events that are regulated during mitosis, including homotypic membrane fusion, spindle pole body function, and ubiquitin-dependent protein degradation. In sperm this intra-acrosomal protein can be used as a marker for evaluation of the physiological state of sperm cells as well as for selection of a suitable method of fertilization in the laboratories of assisted reproduction.,TERA, CDC48