Western blot and Immunohistochemical analyses of RUNX3 expression. Left Panel: Western blot analysis of RUNX3 expression in Jurkat cell lysate. Lysate was prepared from cells of the human Jurkat (Acute T cell leukemia, ATCC TIB-152) cell line. Cell lysate proteins were resolved by electrophoresis (SDS-PAGE) in a 4-20% Tris-Glycine polyacrylamide gel, transferred to PVDF membranes and then probed with 1 µg/ml of Purified Anti-RUNX3 antibody (Cat. No. 564813) followed by HRP-conjugated Goat Anti-Mouse IgG (Cat. No. 554002), and ECL Western blot detection reagents. RUNX3 was identified as protein bands of approximately 40 to 50 kDa. Right Panel: Immunohistochemical staining of RUNX3 expressed in human tonsillar cells. Following antigen retrieval with BD Retrievagen A Buffer (Cat. No. 550524), the formalin-fixed paraffin-embedded tonsil sections were stained with either Purified Mouse IgG1 κ Isotype Control (Cat. No. 550878; Left Image) or Purified Mouse Anti-RUNX3 antibody (Cat. No.564813; Right Image). A three-step staining procedure that employs a Biotin Goat Anti-Mouse Immunoglobulin (Cat. No. 550337), Streptavidin-Horseradish Peroxidase (HRP) (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. As shown in the Right Image, the RUNX3-specific antibody primarily stained the nuclei of some tonsillar lymphocytes. Original magnification: 40×.
Western blot and Immunohistochemical analyses of RUNX3 expression. Left Panel: Western blot analysis of RUNX3 expression in Jurkat cell lysate. Lysate was prepared from cells of the human Jurkat (Acute T cell leukemia, ATCC TIB-152) cell line. Cell lysate proteins were resolved by electrophoresis (SDS-PAGE) in a 4-20% Tris-Glycine polyacrylamide gel, transferred to PVDF membranes and then probed with 1 µg/ml of Purified Anti-RUNX3 antibody (Cat. No. 564813) followed by HRP-conjugated Goat Anti-Mouse IgG (Cat. No. 554002), and ECL Western blot detection reagents. RUNX3 was identified as protein bands of approximately 40 to 50 kDa. Right Panel: Immunohistochemical staining of RUNX3 expressed in human tonsillar cells. Following antigen retrieval with BD Retrievagen A Buffer (Cat. No. 550524), the formalin-fixed paraffin-embedded tonsil sections were stained with either Purified Mouse IgG1 κ Isotype Control (Cat. No. 550878; Left Image) or Purified Mouse Anti-RUNX3 antibody (Cat. No.564813; Right Image). A three-step staining procedure that employs a Biotin Goat Anti-Mouse Immunoglobulin (Cat. No. 550337), Streptavidin-Horseradish Peroxidase (HRP) (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. As shown in the Right Image, the RUNX3-specific antibody primarily stained the nuclei of some tonsillar lymphocytes. Original magnification: 40×.
Product Details
BD Pharmingen™
AML2; AML-2; CBFA3; PEBP2aC; PEBP2A3
Human (QC Testing), Mouse (Tested in Development)
Mouse BALB/c IgG1, κ
Human RUNX3 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested), Immunohistochemistry-paraffin, Western blot (Tested During Development)
40-50 kDa
0.5 mg/ml
AB_2738968
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- An isotype control should be used at the same concentration as the antibody of interest.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
Companion Products
Retrievagen A (pH 6.0) RUO
Cat No.
550524
Purified Mouse IgG1 κ Isotype Control RUO
Size
1 mL
Cat No.
550878
DAB Substrate Kit RUO
Size
500 Tests
Cat No.
550880
HRP Goat Anti-Mouse Ig RUO
Size
1 mL
Cat No.
554002
Biotin Goat Anti-Mouse Ig (Multiple Adsorption) RUO
Size
0.25 mg
Cat No.
550337
Streptavidin HRP RUO
Size
50 mL
Cat No.
550946
564813 Rev. 1
Antibody Details
R3-5G4
The R3-5G4 monoclonal antibody specifically binds to Runt-related transcription factor 3 (RUNX3) which is also known as Acute myeloid leukemia 2 protein (AML2), Core-binding factor subunit alpha-3 (CBFA3), and Polyomavirus enhancer-binding protein 2 alpha C subunit (PEBP2aC). RUNX3 is a member of the RUNX transcription factor family which includes RUNX1-3. RUNX3 is a key regulator of gene expression related to the development and differentiation of cells within the nervous and immune systems. In the immune system, RUNX3 is particularly involved in the commitment of CD8+ T cells in the thymus. RUNX3 is highly expressed and essential for the cytotoxic functions of NK cells, peripheral CD8+T cells and CD4+CD8αα intraepithelial lymphocytes in the gut. RUNX3 also plays a role in the differentiation and effector functions of Th1 cells. RUNX3 is activated downstream of the TGF-β signaling pathway and can play a role in tumor suppression. Aberrant expression of RUNX3 has been associated with tumorigenesis including the development of gastric and other cancers.
564813 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
564813 Rev.1
Citations & References
Development References (6)
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Chuang LS, Ito K, Ito Y. RUNX family: Regulation and diversification of roles through interacting proteins. Int J Cancer. 2013; 132(6):1260-1271. (Biology). View Reference
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Djuretic IM, Cruz-Guilloty F, Rao A. Regulation of gene expression in peripheral T cells by Runx transcription factors. Adv Immunol. 2009; 104(1):23. (Biology). View Reference
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Ito K, Liu Q, Salto-Tellez M, et al. RUNX3, a novel tumor suppressor, is frequently inactivated in gastric cancer by protein mislocalization. Cancer Res. 2005; 65(17):7743-7750. (Immunogen: Western blot). View Reference
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Lotem J, Levanon D, Negreanu V, Leshkowitz D, Friedlander G, Groner Y. Runx3-mediated transcriptional program in cytotoxic lymphocytes. PLoS ONE. 2013; 8(11):e80467. (Biology). View Reference
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Reis BS, Rogoz A, Costa-Pinto FA, Taniuchi I, Mucida D. Mutual expression of the transcription factors Runx3 and ThPOK regulates intestinal CD4(+) T cell immunity. Nat Immunol. 2013; 14(3):271-280. (Biology). View Reference
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Setoguchi R, Tachibana M, Naoe Y, et al. Repression of the transcription factor Th-POK by Runx complexes in cytotoxic T cell devel. Science. 2008; 19(5864):822-825. (Biology). View Reference
564813 Rev. 1
Please refer to Support Documents for Quality Certificates
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.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
