RUNX3 antibody (AA 31-130)

Catalog No. ABIN1714069

This independently validated Rabbit Polyclonal antibody specifically detects RUNX3 in WB, ELISA, IF (p), FACS, IF (cc), IHC (p), ICC and IHC (fro). It exhibits reactivity toward Human.

Quick Overview for RUNX3 antibody (AA 31-130) (ABIN1714069)

Target: RUNX3 (Runt-Related Transcription Factor 3 (RUNX3))

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This RUNX3 antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Flow Cytometry (FACS), Immunofluorescence (Cultured Cells) (IF (cc)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunocytochemistry (ICC), Immunohistochemistry (Frozen Sections) (IHC (fro))

Product Details anti-RUNX3 Antibody

Binding Specificity: AA 31-130

Cross-Reactivity: Human

Predicted Reactivity: Mouse,Rat,Dog,Cow,Sheep,Horse,Chicken

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human Runx3

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200
ICC 1:100-500

Restrictions: For Research Use only

Independent Validation (IF)

Validation #029621 (Immunofluorescence)

by: CaresBio Laboratory

No.: #029621

Date: 03/08/2014

Lot Number: 131016

Method validated: Immunofluorescence

Positive Control: A431 cells

Negative Control: HEK293 cells

Notes: Strong signal was detected in the positive control, and no signal was detected in the negative control.

Validation Images

Figure 1: A431 cells stained with anti-RUNX3 (green) and counterstained with DAPI (blue).

Figure 2: HEK293 cells stained with anti-RUNX3 (green) and counterstained with DAPI (blue).

Figure 3: A431 cells stained with isotype control (green) and counterstained with DAPI (blue).

Figure 4: A431 cells stained with secondary antibody only (green) and counterstained with DAPI (blue).

Full Methods

Primary Antibody:

• Antigen: Runt-Related Transcription Factor 3 (RUNX3)
• Catalog number: ABIN1714069
• Lot number: 131016

Secondary Antibody:

• Antibody: Donkey Anti-Rabbit secondary antibody IgG (H+L) AlexaFluor 488

Full Protocol:

• Antibody staining:
• A431 and Hek-293 cell lines were grown directly on coverslips and fixed with 4% formaldehyde in PBS for 15 min at room temperature (RT).
• Fixed cells were rinsed three times in PBS for 5 min each at RT.
• Cells were blocked in 1X PBS/1% BSA/0.3% Triton™ X-100 to block unspecific binding of the antibodies for 60 min at RT.
• Cells were incubated with primary antibody diluted 1:100 in 1X PBS/1% BSA/0.3% Triton™ X-100 overnight at 4°C.
• Cells were rinsed three times in PBS for 5 min each at RT.
• Cells were incubated with donkey anti rabbit FITC conjugated secondary antibody for 60 min in dark at RT.
• Cells were rinsed three times in PBS for 5 min each at RT.
• Coverslips were mounted on slides with ProLong® Gold Antifade Reagent with DAPI.
• Stained cells were imaged with an Olympus IX73 microscope.
• Isotype Control Staining:
  
• All the steps are done same as previously described until cells were blocked in 1X PBS/1% BSA/0.3% Triton™ X-100 to block unspecific binding of the antibodies for 60 min at RT. - Cells were incubated with 10% normal rabbit serum overnight at 4°C.
• Cells were rinsed three times in PBS for 5 min each at RT.
• Cells were incubated with donkey anti rabbit FITC conjugated secondary antibody for 60 min in dark at RT.
• Cells were rinsed three times in PBS for 5 min each at RT.
• Coverslips were mounted on slides with ProLong® Gold Antifade Reagent with DAPI.
• Stained cells were imaged with an Olympus IX73 microscope.
• Secondary Antibody Only Control Staining:
  
• All the steps are done same as previously described until cells were blocked in 1X PBS/1% BSA/0.3% Triton™ X-100 overnight at 4°C. - Cells were incubated with donkey anti rabbit Alexa488 conjugated secondary antibody for 60 min in dark at RT.
• Cells were rinsed three times in PBS for 5 min each at RT.
• Coverslips were mounted on slides with ProLong® Gold Antifade Reagent with DAPI.
• Stained cells were imaged with an Olympus IX73 microscope.

Experimental Notes:

• No discrepancies reported.
• Runx3 antibody is recommended for IF on A431 cells according to this protocol.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

Precaution of Use: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

Target Details for RUNX3

Target: RUNX3 (Runt-Related Transcription Factor 3 (RUNX3))

Alternative Name: Runx3

Background:

Synonyms: RUNX 3, RUNX-3, Runt-related transcription factor 3, Core-binding factor, alpha 3 subunit, CBF-alpha 3, Acute myeloid leukemia 2 protein, OncogeneAML-2, Polyomavirus enhancer-binding protein 2 alpha C subunit, PEBP2-alpha C, PEA2-alpha C, SL3-3 enhancer factor 1 alpha C subunit, SL3/AKV core-binding factor alpha C subunit, Core-binding factor, alpha 3 subunit, CBF-alpha 3, Acute myeloid leukemia 2 protein, Oncogene AML-2, Polyomavirus enhancer-binding protein 2 alpha C subunit, PEBP2-alpha C, PEA2-alpha C, SL3-3 enhancer factor 1 alpha C subunit, SL3/AKV core-binding factor alpha C subunit.

Background: RUNX3 binds to the core site of murine Leukemia virus, the core sequences in the enhancer of the polyomavirus, and also to the enhancers of the T-cell receptor genes. May be involved in the control of cellular proliferation and/or differentiation (By similarity). Heterodimer of an alpha and a beta subunit. The alpha subunit binds DNA as a monomer and through the Runt domain.DNA-binding is increased by heterodimerization. Interacts with TLE1 and SUV39H1, Subcellular location in Nucleus.

Gene ID: 864