Human vWF-A2 Antibody

Catalog #: MAB11543 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB11543-100
MAB11543-SP

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VWF in Human Non-Hodgkins Lymphoma via seqIF™ staining on COMET™

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Human vWF-A2 Antibody Summary

Species Reactivity

Human

Specificity

Detects recombinant human VWF in Direct ELISA.

Source

Monoclonal Mouse IgG_2B Clone # 1072103

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human VWF
Asp1498-Val1665
Accession # P04275

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Immunohistochemistry

3-25 µg/mL

Immersion fixed paraffin-embedded sections of human placenta

Multiplex Immunofluorescence

1 µg/mL

Immersion fixed paraffin-embedded sections of human non-Hodgkin's Lymphoma

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

Multiplex Immunofluorescence

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VWF in Human Non-Hodgkins Lymphoma via seqIF™ staining on COMET™ VWF was detected in immersion fixed paraffin-embedded sections of human non-Hodgkin's Lymphoma using Mouse Anti-Human VWF, Monoclonal Antibody (Catalog #MAB11543) at 1ug/mL at 37° Celsius for 4 minutes. Before incubation with the primary antibody, tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing using PreTreatment Module (PT Module) and Dewax and HIER Buffer H (pH 9; Epredia Catalog # TA-999-DHBH). Tissue was stained using the Alexa Fluor™ 647 Goat anti-Mouse IgG Secondary Antibody at 1:200 at 37 ° Celsius for 2 minutes. (Yellow; Lunaphore Catalog # DR647MS) and counterstained with DAPI (blue; Lunaphore Catalog # DR100). Specific staining was localized to the cytoplasm of endothelial cells. Protocol available in COMET™ Panel Builder.

Immunohistochemistry

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Detection of vWF‑A2 in Human Placenta. vWF‑A2 was detected in immersion fixed paraffin-embedded sections of human placenta using Mouse Anti-Human vWF‑A2 Monoclonal Antibody (Catalog # MAB11543) at 25 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001) or the HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm of endothelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute lyophilized material at 0.2mg/ml in sterile PBS. For liquid material, refer to CoA for concentration.

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: vWF-A2

von Willebrand Factor (vWF) is a large, multimeric glycoprotein made by endothelial cells and megakaryocytes. The pre-pro-vWF protein contains 2813 amino acids (aa), which consists of 22 aa signal peptide, 741 aa propeptide, and mature vWF monomer of 2050 aa (1-4). The pro-vWF undergoes dimerization in the endoplasmic reticulum (ER) through C-terminal “cysteine-knot” (CK) domain. The pro-vWF dimmers are transported to Golgi and form multimers by forming disulfide bond in amino-terminal region of the mature form. The proteolytic processing of pro-region also occurs in Golgi. The matured vWF is stored in Weibel-Pallade bodies in endothelial cells and granules in megakaryocytes and platelets. The unusually-large vWF (ulvWF) multimers released from cells are very efficient in binding to platelets to form thrombus. The population of these highly active ulvWF multimers is controlled by a specific protease, ADAMTS13, which cleaves between residues Tyr1605 and Met1606 in the A2 domain of vWF. In the plasma, vWF appears as a series of large and intermediate multimers with molecular masses from several thousand to 500 kDa. vWF also performs hemostatic functions (3-5). In a high shear-stressed environment, vWF undergoes conformational change to expose a binding site for glycoprotein Ib alpha. As a result, vWF facilitates aggregation of platelets. In addition to platelet binding, vWF binds coagulation factor VIII to increase the lifetime of FVIII in plasma. The purified rhvWF-A2 contains the A2 domain of vWF.

References