Ephrin B1, B2, B3 Antibody

SKU: orb614006

Description

Ephrin B1-B3 antibody

Images & Validation

• Tested Applications: ELISA, IF, WB
• Dilution Range: ELISA: 1:10,000-50,000, IF: 1:5000, WB: 1:1000
• Reactivity: Human, Mouse, Xenopus
• Application Notes:
  Anti-Ephrin B1, B2, B3 Antibody has been tested in ELISA, Western Blot, and Immunofluorescence. Expect a band at ~35-38 kDa in western blot using appropriate lysates or tissues. Positive control used Xenopus laevis and mouse kidney in WB and Xenopus laevis in IF.

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Immunogen: Anti-Ephrin B1, B2, B3 antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to C-terminal portion of human Ephrin B1, B2, B3 conjugated to Keyhole Limpet Hemocyanin (KLH).
• Purity: This affinity purified antibody is directed against human Ephrin B1, B2, B3. This product was affinity purified from monospecific antiserum by immunoaffinity purification. A BLAST analysis was used to suggest cross-reactivity with the antigen based on 100% homology with Ephrin-B1 to human, mouse, rat, and Gallus gallus; 100% homology with Ephrin-B2 to mouse; 100% homology with Ephrin-B3 to human and mouse.
• Conjugation: Unconjugated

Storage & Handling

• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Form/Appearance: Liquid (sterile filtered)
• Buffer/Preservatives: Preservative: 0.01% (w/v) Sodium Azide. Stabilizer: None; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
• Concentration: 1.05 mg/mL
• Expiration Date: 12 months from date of receipt.
• Dry Ice Shipping: Please note: This product requires shipment on dry ice. A dry ice surcharge will apply.
• Disclaimer: For research use only

Alternative Names

Rabbit Anti-Ephrin B1, B2, B3 Antibody, Ephrin-B1, Ephrin-B2, Ephrin-B3, ELK Ligand, EPLG2, LERK2, EFL3, Craniofrontonasal Syndrome (Craniofrontonasal Dysplasia), EPH-Related Receptor Transmembrane Ligand ELK-L3, Eph-Related Receptor Tyrosine Kinase Ligand 2, Eph-Related Receptor Tyrosine Kinase Ligand 8, EPH-Related Receptor Tyrosine Kinase Ligand 2, LERK-2, LERK-8, Elk-L, EFL-3, ELK-L, CFND, EFL6, EFB1, CFNS

Immunofluorescence of Rabbit Anti-Ephrin B1-B3 Antibody. (A). Neural Plate of Xenopus laevis stage 17. (B). Knockdown neural plate of Xenopus laevis stage 17. Primary Antibody: Anti-Ephrin B1-3 at 1:5000 overnight at 2-8°C. Secondary Antibody: Anti-Rabbit IgG Fluorescent Secondary. Fixative: 4% Paraformaldehyde in PBS. Permeabilization: 0.05% Triton-X100 in PBS. Nuclear Counterstain: None. Magnification: 63X objective, Zeiss LSM710. Results: Ephrin B1 B2 B3 triple knockdown with a mixture of all three morpholinos abolished most of the signal which can be seen at expected plasma membrane and vesicle subcellular locations.

Western Blot of Rabbit Anti-Ephrin B1-B3 Antibody. Lane 1: 1st experiment: Uninjected heterogenous embryo [+]. Lane 2: 1st experiment: Ephrin B1, 2 MO (2 morpholino oligomers). Lane 3: 2nd experiment: Ephrin B1, 2, 3 MO (3 morpholino oligomers). Lane 4: 2nd experiment: Ephrin B3 MO (1 morpholino oligomer). Lane 5: 2nd experiment: Uninjected heterogenous embryo [+]. Primary Antibody: Anti-Ephrin B1-B3 at 1:2000 overnight at 2-8°C. Secondary Antibody: Goat anti-Rabbit IgG HRP (p/n orb347654) 1:5000 for 60 min at RT. Block: 5% skim milk/TBST/0.1% Tween. Exposure: 10 sec. Predicted MW: ~38kDa. Observed: ~37-45kDa. Results: Ephrin B1, 2, 3 triple knockdown with a mixture of all three morpholinos abolished significant amount of the WB signal at expected size. The comparison of lane 3 and 4 suggests not much endogenous Ephrin B3 expression exist in this stage embryos.

Western Blot of Rabbit Anti-Ephrin B1-B3 Antibody. Lane 1: Opal PreStain Molecular Weight Marker. Lane 2: EPHRIN B1 overexpressing HEK293T Lysate (10 µg) [+]. Lane 3: HEK293T Lysate (10 µg) [-]. Lane 4: Mouse Kidney Lysate (p/n orb348717) (35 µg) [+]. Primary Antibody: Anti-Ephrin B1-B3 at 1:1000 overnight at 2-8°C. Secondary Antibody: Goat anti-Rabbit IgG MX10 HRP (p/n orb347654) at 1:70000 for 30 min at RT. Block: BlockOut Buffer (p/n orb348644). Predicted MW: ~38kDa endogenous, ~37-45kDa due to glycosylation. Observed: ~46-52kDa overexpressed, ~34, ~40 kDa endogenous.

Western Blot of Rabbit Anti-Ephrin B1-B3 Antibody. Lane 1: Uninjected Xenopus laevis heterogenous embryo control (20 µl). Lane 2: Morpholino Knockdown of Ephrin B1/2 heterogenous embryo (20 µl). Lane 3: Morpholino Knockdown of Ephrin B1/2 heterogenous embryo (20 µl). Lane 4: Ephrin B1-HA overexpression heterogenous embryo lysate (20 µl). Lane 5: Ephrin B2-HA overexpression heterogenous embryo lysate (20 µl). Primary Antibody: Anti-Ephrin B1-B3 at 1:2000 overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG HRP (p/n orb347654) at 1:5000 for 60 mins at RT. Block: 5% skim milk/TBST/0.1% Tween for 30 mins at RT. Predicted MW: ~38kDa. Observed MW: ~37-45kDa (due to glycosylation). Exposure: 10 sec. Results: Ephrin B1, 2, double knockdown with a mixture of both morpholinos oligomers abolished most of the WB signal at expected size which was double checked by over-expression.

Quick Database Links

UniProt

P52800

RefSeq (Protein):NP_004420.1