Fluorescent TrueBlot®: Anti-Rabbit IgG DyLight™ 680

Details for Product No. ABIN6698839
Target Name (Antigen)
Reactivity
Rabbit
Host
Mouse
Conjugate
DyLight 680
Application
FLISA, Flow Cytometry (FACS), Fluorescence Microscopy (FM), Immunohistochemistry (IHC), Western Blotting (WB)
Options
Brand TrueBlot®
Characteristics

Synonyms: DL680, TrueBlot, DL680 TrueBlot ULTRA, DyLight™ 680 TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting, Fluorescent TrueBlot, Rb TrueBlot

Background: Rabbit IgG TrueBlot® is a unique DyLight™ 680 conjugated Anti-rabbit IgG immunoblotting (second step) reagent. Rabbit IgG TrueBlot® enables detection of immunoblotted target protein bands, without hindrance by interfering immunoprecipitating immunoglobulin heavy and light chains. It is easy to generate publication-quality IP/Fluorescent Western Blot data with Rabbit IgG TrueBlot®, simply substitute the conventional DL680 Anti-rabbit IgG blotting reagent with Fluorescent Rabbit TrueBlot® Antibody DyLight™ 680 and follow the prescribed protocol for sample preparation and immunoblotting. Rabbit IgG TrueBlot® is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot preferentially detects the non-reduced form of rabbit IgG over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Rabbit IgG TrueBlot® with the 55 kDa heavy chains and the 23 kDa light chains of the immunoprecipitating antibody is minimized thereby eliminating interference by the heavy and light chains of the immunoprecipitating antibody in IP/Western blot applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.

Purification Fluorescent Rabbit TrueBlot® Antibody DyLight™ 680 Conjugate was prepared from tissue culture supernatant by Protein G affinity chromatography. Assay by Immunoelectrophoresis resulted in a single precipitin arc against Anti-Rabbit Serum. Reactivity is observed against native Rabbit IgG by both Western blot and ELISA.
Labeling Ratio 2.3
Target Name (Antigen)
Application Notes

Immunohistochemistry Dilution: User Optimized

Application Note: Fluorescent Rabbit TrueBlot® Antibody DyLight™ 680 may also be used for detection in immunoassays that do not employ immunoprecipitation. Fluorescent Rabbit TrueBlot® Antibody DyLight™ 680 is provided as a lyophilized powder. To conserve reagent, we recommend incubating the blots from minigels in sealed bags (removing as much air as possible) with minimal volume (2-3 mLs). If used conservatively at 2.5 mLs/blot will yield enough reagent for 40 blots. Note that there are three key procedural considerations: 1. Protein A or G should not be used for the immunoprecipitation. Use of protein A or G beads with the rabbit TrueBlot will result in contaminating bands. For immunoprecipitation, anti-rat IgG beads, or anti-rabbit IgG beads should be used for rat or rabbit immunoprecipitating antibodies, respectively. 2. Immunoprecipitate should be completely reduced. 3. MB-070 Blocking Buffer for Fluorescent Western Blotting should be used as the blocking protein for the immunoblot. All recommended dilutions for listed applications are intended as an initial recommendation, specific conditions for each protein and antibody combination should be specifically optimized by the end user.

FLISA Dilution: User Optimized

Flow Cytometry Dilution: 1:2,000 - 1:10,000

Western Blot Dilution: 1:1000

IF Microscopy Dilution: 1:500 - 1:2,500

Restrictions For Research Use only
Format Lyophilized
Reconstitution

Reconstitution Volume: 100 μL

Reconstitution Buffer: Restore with deionized water (or equivalent)

Buffer

Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Stabilizer: 10 mg/mL Polyethylene Glycol (PEG-8000)

Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage RT,4 °C,-20 °C
Storage Comment Store vial at 4 °C prior to restoration. For extended storage aliquot contents and freeze at -20 °C or below. 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.
Expiry Date 12 months
Supplier Images
Image no. 1 for Fluorescent TrueBlot®: Anti-Rabbit IgG DyLight™ 680 (ABIN6698839) Rabbit IP / Western Blot: Jurkat cell lysate (0.5 ml of 1x10e7 cells/ml) was incubate...
Image no. 2 for Fluorescent TrueBlot®: Anti-Rabbit IgG DyLight™ 680 (ABIN6698839) Fluorescent: Anti-Rabbit IgG DyLight 680 Conjugated - Western Blot. Western Blot of F...
Product cited in: Tian, Li, Gao, Li, Yang, Wang: "Identification and validation of the role of matrix metalloproteinase-1 in cervical cancer." in: International journal of oncology, Vol. 52, Issue 4, pp. 1198-1208, 2018 (PubMed).

Zu, Wang, Ping, Sun: "Tan IIA inhibits H1299 cell viability through the MDM4‑IAP3 signaling pathway." in: Molecular medicine reports, Vol. 17, Issue 2, pp. 2384-2392, 2018 (PubMed).