Human/Rat Vinculin Antibody

Catalog #: AF6896 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF6896
AF6896-SP

Detection of Human and Rat Vinculin by Western Blot.

3 Images

All Vinculin Antibodies

Product Details

Supplemental Products

Reviews

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

Human/Rat Vinculin Antibody Summary

Species Reactivity

Human, Rat

Specificity

Detects recombinant human Vinculin in direct ELISAs and human and rat Vinculin in Western blots.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human Vinculin
Lys1020-Gln1134
Accession # P18206

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

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

View Larger

Detection of Human and Rat Vinculin by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and rat heart tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Rat Vinculin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6896) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Vinculin at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western

View Larger

Detection of Human Vinculin by Simple Western^TM. Simple Western shows lysates of Exosome Standards (PC‑3) (NBP2-49856) and HepG2 human hepatocellular carcinoma cell line, loaded at 0.5 mg/ml. A specific band was detected for Vinculin at approximately 119 kDa (as indicated) using 10 µg/mL of Sheep Anti-Human/Rat Vinculin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6896). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Immunohistochemistry

View Larger

Vinculin in Human Uterus. Vinculin was detected in immersion fixed paraffin-embedded sections of human uterus using Sheep Anti-Human/Rat Vinculin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6896) at 3 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counter-stained with hematoxylin (blue). vView 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: Vinculin

VCL (Vinculin; from the Latin vinculum denoting a union or connection) is a 115-120 kDa cytoplasmic member of the vinculin/ alpha -catenin family of proteins. The VCL gene is ubiquitously expressed and generates a series of globular domains whose N-and C-terminal globular regions self-associate to generate an inactive molecule. When at least two ligands bind to the molecule, it opens up, allowing for the formation of multiple complexes that impact both signaling and cell structure. Among other things, VCL is known to regulate the expression of E-cadherin, and to connect actin to integrins via talin. Human VCL is 1134 amino acids (aa) in length, and at this size, is commonly known as metavinculin. It contains an 835 aa N-terminal globular head that binds talin, beta - and alpha -catenin, followed by a Pro-rich linker segment (aa 836-878) and a 27-29 kDa C-terminal tail that binds lipids plus paxillin and PKC alpha (aa 879-1134). At least seven utilized phosphorylation sites exist. There are three potential splice variants. One shows a deletion of aa 916-983 and is called vinculin. A second contains an alternative start site at Met74, and again possesses a deletion of aa 916-983. A third shows a 34 aa substitution for aa 262‑1134. Over the range of aa used as the immunogen for this reagent (aa 1020-1134 in metavinculin; 952-1066 in vinculin), human and mouse VCL are identical in aa sequence.

Long Name

Metavinculin

Entrez Gene IDs

7414 (Human); 22330 (Mouse); 305679 (Rat)

Alternate Names

CMD1W; CMH15; metavinculin; MVCL; VCL; Vinculin