RUNX3 antibody (AA 31-130)

Catalog No. ABIN1714069

Product Details anti-RUNX3 Antibody

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

Binding Specificity: AA 31-130

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))

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 (RUNX3 Products)

Synonyms: runxb antibody, RUNX3 antibody, CBFA3 antibody, frrunx3 antibody, runx3 antibody, aml2 antibody, cbfa3 antibody, pebp2ac antibody, AML2 antibody, Cbfa3 antibody, Pebp2a3 antibody, PEBP2aC antibody, runt-related transcription factor 3 antibody, runt related transcription factor 3 antibody, runt related transcription factor 3 L homeolog antibody, runx3 antibody, RUNX3 antibody, runx3.L antibody, Runx3 antibody

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