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Goat anti-Rabbit IgG (H&L), DyLightŪ 488 conjugated

181 €

AS09 633 | clonality: polyclonal  |  host: goat  | reactivity: rabbit IgG (H&L)

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AS09 633

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product information
Background
Goat anti-rabbit IgG, DyLight® 488 Conjugate is a secondary antibody conjugated to DyLight® 488, which binds to all rabbit IgGs in immunological assays.

DyLight® is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
Immunogen

Purified Rabbit IgG, whole molecule

Host Goat
Clonality Polyclonal
Clone
Purity Affinity purified goat IgG
Format Lyophilized
Quantity 1 mg
Reconstitution For reconstitution add 1.1 ml of sterile water. Let it stand 30 minutes at room temperature to dissolve. Prepare fresh working dilutions daily.
Storage
Store lyophilized material at 2-8°C. Product is stable for 4 weeks at 2-8°C after rehydration.
For long time storage after reconstitution, dilute the antibody solution with glycerol to a final concentration of 50% glycerol and store as liquid at -20°C, to prevent loss of enzymatic activity.
For example, if you have reconstituted 1 mg of antibody in 1.1 ml of sterile water add 1.1 ml of glycerol. Such solution will not freeze in -20°C. If you are using a 1:5000 dilution prior to diluting with glycerol, then you would need to use a 1:2500 dilution after adding glycerol. Prepare working dilution prior to use and then discard. Be sure to mix well but without foaming.
Tested applications Immunocytochemistry (ICC), Immunohistochemistry (IHC), Immunofluorescence (IF)
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Plant and algal protein extraction buffer

Additional information
Conjugate is present in 10 mM Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 1 % (w/v) BSA, Protease/IgG free. 0.05 % (w/v) sodium azide is added as preservative.

DyLight® 488 has a maximum absorbance at 493 nm; Emax = 518 nm.
application information
Recommended dilution 1 : 50- 1 : 5 000 (ICC), 1 : 20- 1 : 2000 (IHC), 1 : 3000 (IF)
Expected | apparent MW
Confirmed reactivity Rabbit IgG heavy and light chains (H&L)
Predicted reactivity Rabbit IgG Heavy and Light chains (H&L)
Not reactive in
Additional information
Based in immunoelectrophoresis, this antibody reacts with heavy chains on rabbit IgG and light chains on all rabbit immunoglobulins.

No reactivity is observed to non-immunoglobulin rabbit serum proteins based in immunoelectrophoresis. 
Purity of this antibody is > 95% based on SDS-PAGE.
Selected references Liu et al. (2016). Fold formation at the compartment boundary of Drosophila wing requires Yki signaling to suppress JNK dependent apoptosis. Sci Rep. 2016 Nov 29;6:38003. doi: 10.1038/srep38003.
Wang et al. (2016). Complementary expression of optomotor-blind and the Iroquois complex promotes fold formation to separate wing notum and hinge territories. Dev Biol. 2016 Aug 1;416(1):225-34. doi: 10.1016/j.ydbio.2016.05.020. Epub 2016 May 19
Kaulmann et al. (2016). Inflammation related responses of intestinal cells to plum and cabbage digesta with differential carotenoid and polyphenol profiles following simulated gastro-intestinal digestion. Mol Nutr Food Res. 2016 Mar 18. doi: 10.1002/mnfr.201500947.
Jimenez-Lopez et al. (2015). Biogenesis of protein bodies during legumin accumulation in developing olive (Olea europaea L.) seed. Protoplasma. 2015 May 21.

Application example

immunolocalization of TOPO2 in plant chromosomes

Seeds of field bean (Vicia faba L. subsp. minor var. Nadwiślański; DANKO Group; Sobiejuchy) were sterilized using sodium hypochlorite (0.3% v/v) and germinated in Petri dishes on wetted filter paper at room temperature. At 4 d after imbibition, dark-grown seedlings with primary roots 25±5 mm long were selected for experiments. During incubations roots were oriented horizontally in a humid chamber and aerated continuously on a rotary water-bath shaker (30 rpm) at 23°C. Immunocytochemical assays were performed according to the method prescribed earlier (Rybaczek and Maszewski 2006). Excised apical parts of roots (1.5 mm long) were fixed for 45 min (18°C) in PBS-buffered 3.7% paraformaldehyde, washed several times with PBS and placed in a citric acid-buffered digestion solution (pH 5.0; 37°C for 45 min) containing 2.5% pectinase (Fluka), 2.5% cellulase (Onozuka R-10; Serva) and 2.5% pectoliase (ICN). After removing the digestion solution, root tips were washed 3 times in PBS, rinsed with distilled water and squashed onto Super Frost Plus glass slides (Menzel-Gläser). Air-dried slides were pretreated with PBS-buffered 5% BSA at 20°C for 50 min and incubated overnight in a humidified atmosphere (4°C) with rabbit antibody raised against TOPO2 (Agrisera), dissolved in PBS containing 1% BSA (at a dilution of 1:500). Following incubation, slides were washed 3 times with PBS and incubated for 1 h (18°C) with secondary goat anti-rabbit IgG DyLight®488 antibody (Agrisera, AS09 633, 1:3000). Nuclear DNA was stained with 4’,6-diamidino-2-phenyl-indole (DAPI, 0.4 μg/ml; Sigma-Aldrich). Following washing with PBS, slides were air dried and embedded in Vectashield Mounting Media for Fluorescence (Vector Laboratories). Observations were made using Optiphot-2 fluorescence microscope (Nikon) equipped with B-2A filter (blue light; λ ≈ 495 nm) for DyLight-conjugated antibodies and UV-2A filter (UV light; λ ≈ 365 nm) for DAPI. All images were recorded at exactly the same time of integration using DXM 1200 CCD camera.

Courtesy Dr. Dorota Rybaczek, Lodz University, Poland


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