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Catalog Number | orb614006 |
---|---|
Category | Antibodies |
Description | Ephrin B1-B3 antibody |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, IF, WB |
Reactivity | Human, Mouse, Xenopus |
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). |
Concentration | 1.05mg/mL |
Dilution range | ELISA: 1:10,000-50,000, IF: 1:5000, WB: 1:1000 |
Form/Appearance | Liquid (sterile filtered) |
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 |
UniProt ID | P52800 |
NCBI | NP_004420.1 |
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. |
Buffer/Preservatives | 0.01% (w/v) Sodium Azide |
Alternative names | Rabbit Anti-Ephrin B1, B2, B3 Antibody, Ephrin-B1, Read more... |
Note | For research use only |
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. |
Expiration Date | 12 months from date of receipt. |
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: ~38 kDa. Observed: ~37-45 kDa. 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: ~38 kDa endogenous, ~37-45 kDa due to glycosylation. Observed: ~46-52 kDa 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: ~38 kDa. Observed MW: ~37-45 kDa (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.
ELISA, IF, WB | |
Human, Mouse, Xenopus | |
Rabbit | |
Polyclonal | |
Unconjugated |
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