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Antibodies

Fluorescent TrueBlot[R]: Anti-Mouse Ig DyLight[TM] 680

Product Sizes
100 ul
18-4417-32-100UL
About this Product
SKU:
18-4417-32
Additional Names:
Anti-Mouse IgG DL680, TrueBlot, DL680 TrueBlot ULTRA, DyLight™ 680 TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting, Fluorescent TrueBlot, Ms TrueBlot, IRDye 700, IRDye 680
Application:
ELISA, FLISA, Flow Cytometry, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Western Blot
Buffer:
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride
CE/IVD:
RUO
translate.label.attr.clone:
eB144
Clonality:
Monoclonal
Conjugate:
DyLight™ 680
Concentration:
1 mg/ml
Extra Details:
Mouse IgG TrueBlot® is a unique DyLight™ 680 conjugated Anti-mouse IgG monoclonal secondary antibody. Mouse 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 Mouse IgG TrueBlot®, simply substitute the conventional DL680 Anti-mouse IgG blotting reagent with Fluorescent Mouse TrueBlot® Antibody DyLight™ 680 and follow the prescribed protocol for sample preparation and immunoblotting. Mouse IgG TrueBlot® is ideal for use in protocols involving immunoblotting of immunoprecipitated proteins. TrueBlot preferentially detects the non-reduced form of mouse IgG over the reduced, SDS-denatured form of IgG. When the immunoprecipitate is fully reduced immediately prior to SDS-gel electrophoresis, reactivity of Mouse 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 blotting applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.
Formulation:
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Host:
Rat
Isotype:
IgG
Physical State:
Lyophilized
Reactivities:
Mouse
Shipping Conditions:
Ambient
Specificity:
Fluorescent Mouse 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-Mouse Serum. Reactivity is observed against native Mouse IgG by both Western blot and ELISA.
Storage Conditions:
Please refer to datasheet
Supplier:
Rockland Inc
Type:
Antibody: Secondary Antibodies
Mouse TrueBlot® IP / Western Blot: Caspase 7 was immunoprecipitated from 0.5 ml of 1x10e7 Jurkat cells/ml with 5 ug mouse anti-human Caspase 7. Precipitate from 1x10e6 cells was subjected to electrophoresis, transferred to an PVDF membrane, and Western blotted with anti-Caspase 7 using Mouse TrueBlot® ULTRA: Anti-Mouse Ig HRP (Lane 1) or conventional HRP-conjugated anti-mouse antibody (Lane 2) - note the detection of the heavy and light chains of the immunoprecipitating antibody in Lane 2 but not in Lane 1. When Lane 1 is re-immunoblotted using conventional HRP-conjugated anti-mouse polyclonal antibody (Lane 3), the heavy and light chains are now detected, confirming that although the immunoprecipitating heavy and light chains are present, Mouse TrueBlot® ULTRA: Anti-Mouse Ig HRP detects only native antibody and not denatured heavy and light chains.
Western Blot of Fluorescent TrueBlot®: Anti-Mouse Ig DyLight 680 Conjugated. Lane 1: Mouse IgG, Non-reduced. Lane 2: Mouse IgG, Reduced. Load: 50 ng per lane. Primary antibody: none. Secondary antibody: Fluorescent TrueBlot®: Anti-Mouse Ig DyLight 680 Conjugated at 1:1,000 for 60 min at RT. Block: MB-070 for 30 min at RT. Predicted/Observed size: 160 kDa for Mouse IgG, Non-reduced. Migrates at slightly higher molecular weight. Other band(s): none.
Immunofluorescence microscopy of a-tubulin in A431 cells using DyLight™ 680-conjugated Fluorescent TrueBlot® anti-mouse IgG for detection. A431 cells were fixed with 100% methanol, blocked (5% rat serum/0.3% Triton X-100 in 1X PBS) for 1 hr, then incubated with 15 ug/mL of anti-a-tubulin primary antibody (Cat. No. 200-301-880) at 4°C overnight. After 3 washes in 1X PBS for 5 min each, 5 ug/mL of Fluorescent TrueBlot® anti-mouse IgG DyLight™ 680 was added and allowed to incubate for 1 hr at room temperature. Nuclei were counterstained with DAPI present in mounting medium. The predicted main localization is microtubules. Image taken at 63X magnification. (a) Merged a-tubulin (red)/DAPI (blue) image shown. (b) secondary antibody only.
Immunofluorescence microscopy of A Alpha-tubulin in HeLa cells using DyLight™ 680-conjugated Fluorescent TrueBlot® anti-mouse IgG (p/n 18-4417-32) for detection. HeLa cells were fixed with 100% methanol, blocked (5% rat serum/0.3% Triton X-100 in 1X PBS) for 1hr, then incubated with 15ug/mL of anti-alpha-tubulin primary antibody (p/n 200-301-880) at 4°C overnight. Following 3 washes in 1X PBS for 5 min each, 5ug/mL of Fluorescent TrueBlot® anti-mouse IgG DyLight™ 680 was added and allowed to incubate for 1hr at room temperature. Nuclei were counterstained with DAPI present in mounting medium. The predicted main localization is microtubules. Image taken at 63X magnification. (a) Merged a-tubulin (red)/DAPI (blue) image shown. (b) secondary antibody only.