BD Pharmingen™ Purified Human anti-VASA/DDX4
Clone R11-A3
Western Blot analysis of VASA/DDX4 in mouse embryonic stem (ES) cell line. Lysate from ES-E14TG2a cells (ATCC CRL-1821) was probed with Purified Human anti-VASA/DDX4 Fab at titrations of 0.25 (lanes 1), 0.125 (lanes 2), and 0.062 µg/ml (lanes 3). VASA/DDX4 is identified as a band of 76 kDa in the mouse ES cells.
Western Blot validation of VASA/DDX4 by RNAi in mouse ES cell line. Lysates from ES-E14TG2a cells (24 hour, lane 1, and 48 hour, lane 2) and VASA/DDX4 RNAi-transfected ES-E14TG2a cells (24 hour, lane 2, and 48 hour, lane 4) were probed with Purified Human anti-VASA/DDX4 Fab at 1.0 µg/ml. Down-regulation of VASA/DDX4 expression is evident in the RNAi-transfected cells. Purified Mouse anti-Actin monoclonal antibody (Cat. No. 612656 or 612657, 1:2000) was the gel-loading control.
VASA/DDX4 staining on human (left column) and mouse (right column) testes. Following antigen retrieval with BD Retrievagen A buffer (Cat. no. 550524), the formalin-fixed paraffin-embedded sections were stained with either Purified Human anti-VASA/DDX4 Fab (top row) or a control Fab specific for an irrelevant antigen (bottom row), with Hematoxylin counterstaining. VASA/DDX4 is detected in spermatids and spermatocytes in the seminiferous tubules. Original magnification: 20X.
Western Blot analysis of VASA/DDX4 in mouse embryonic stem (ES) cell line. Lysate from ES-E14TG2a cells (ATCC CRL-1821) was probed with Purified Human anti-VASA/DDX4 Fab at titrations of 0.25 (lanes 1), 0.125 (lanes 2), and 0.062 µg/ml (lanes 3). VASA/DDX4 is identified as a band of 76 kDa in the mouse ES cells.
Western Blot validation of VASA/DDX4 by RNAi in mouse ES cell line. Lysates from ES-E14TG2a cells (24 hour, lane 1, and 48 hour, lane 2) and VASA/DDX4 RNAi-transfected ES-E14TG2a cells (24 hour, lane 2, and 48 hour, lane 4) were probed with Purified Human anti-VASA/DDX4 Fab at 1.0 µg/ml. Down-regulation of VASA/DDX4 expression is evident in the RNAi-transfected cells. Purified Mouse anti-Actin monoclonal antibody (Cat. No. 612656 or 612657, 1:2000) was the gel-loading control.
VASA/DDX4 staining on human (left column) and mouse (right column) testes. Following antigen retrieval with BD Retrievagen A buffer (Cat. no. 550524), the formalin-fixed paraffin-embedded sections were stained with either Purified Human anti-VASA/DDX4 Fab (top row) or a control Fab specific for an irrelevant antigen (bottom row), with Hematoxylin counterstaining. VASA/DDX4 is detected in spermatids and spermatocytes in the seminiferous tubules. Original magnification: 20X.
Product Details
BD Pharmingen™
DDX4
Mouse (QC Testing), Human (Reactivity Confirmed in Development)
Human Ig, λ
immunogen is NA for phage display technology
Western blot (Routinely Tested), Immunohistochemistry-formalin (antigen retrieval required) (Tested During Development)
76 kDa
0.5 mg/ml
AB_1645336
Aqueous buffered solution containing ≤0.09% sodium azide.
Preparation And Storage
The recombinant Fab was purified from tissue culture supernatant by affinity chromatography. Store undiluted at 4°C.
Recommended Assay Procedures
We recommend Monoclonal ANTI-FLAG® M2-Peroxidase (Sigma-Aldrich, Cat. No. A8592) for use as a second-step reagent for western blots and immunohistochemistry.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
560189 Rev. 1
Antibody Details
R11-A3
The R11-A3 Fab reacts with a protein of the Asp-Glu-Ala-Asp (DEAD) box (DDX) family that is homologous to the VASA protein of Drosophila. The VASA/DDX4 gene is conserved in invertebrate and vertebrate species, and its expression is germ cell specific. In the human, VASA/DDX4 is expressed in fetal and adult germ cells, both male and female, but not in other fetal and adult tissues. Studies of VASA/DDX4 expression in various types of tumors suggest that expression of VASA/DDX4 is a useful biomarker for germ cell cancers.
The R11-A3 Fab was selected by screening a human Fab phage display library (Hoet et al.) licensed from Dyax Corp (Cambridge, MA) with immobilized recombinant human VASA/DDX4 protein.
560189 Rev. 1
Format Details
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
560189 Rev.1
Citations & References
View product citations for antibody "560189" on CiteAb
Development References (6)
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Abdelhaleem M. RNA helicases: regulators of differentiation. Clin Biochem. 2005; 38:499-503. (Biology: Western blot, Immunohistochemistry-formalin (antigen retrieval required)).
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Castrillon DH, Quade BJ, Wang TY, Quigley C, Crum CP. The human VASA gene is specifically expressed in the germ cell lineage. Proc Natl Acad Sci U S A. 2000; 97(17):9585-9590. (Biology: Western blot, Immunohistochemistry-formalin (antigen retrieval required)).
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Chu DS, Liu H, Nix P, et al. Sperm chromatin proteomics identifies evolutionarily conserved fertility factors. Nature. 2006; 443:101-105. (Biology: Western blot, Immunohistochemistry-formalin (antigen retrieval required)).
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Clark AT, Bodnar MS, Fox M, et al. Spontaneous differentiation of germ cells from human embryonic stem cell in vitro. Hum Mol Genet. 2004; 13(7):727-739. (Biology: Western blot, Immunohistochemistry-formalin (antigen retrieval required)).
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Hoet RM, Cohen EH, Kent RB, et al. Generation of high-affinity human antibodies by combining donor-derived and synthetic complementarity-determining-region diversity. Nat Biotechnol. 2005; 23(3):344-348. (Methodology: Western blot, Immunohistochemistry-formalin (antigen retrieval required)). View Reference
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Zeeman AM, Stoop H, Boter M, et al. VASA is a specific marker for both normal and malignant human germ cells. Lab Invest. 2002; 82(2):159-166. (Biology: Western blot, Immunohistochemistry-formalin (antigen retrieval required)). View Reference
560189 Rev. 1
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
