Fluorescent TrueBlot®: Anti-Mouse Ig Fluorescein

Details for Product No. ABIN6698838
Target Name (Antigen)
  • ATPIG
  • ATPIQ
  • A330005H02Rik
  • AI315324
  • Ig
  • ATPase phospholipid transporting 11C
  • ATPase, class VI, type 11C
  • ATP11C
  • Atp11c
Reactivity
Mouse (Murine)
Host
Rat
Conjugate
FITC
Application
FLISA, Flow Cytometry (FACS), Fluorescence Microscopy (FM), Immunohistochemistry (IHC), Western Blotting (WB)
Options
Brand TrueBlot®
Characteristics

Synonyms: FITC, TrueBlot, FITC TrueBlot ULTRA, Fluorescein TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting, Fluorescent TrueBlot, Ms TrueBlot

Background: Mouse IgG TrueBlot® is a unique fluorescein conjugated Anti-mouse IgG immunoblotting (second step) reagent. 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 FITC Anti-mouse IgG blotting reagent with Fluorescent Mouse TrueBlot® Antibody Fluorescein 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 blot applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.

Purification Fluorescent Mouse TrueBlot® Antibody Fluorescein Conjugate was prepared from tissue culture supernatant by Protein G affinity chromatography. Assay by Immunoelectrophoresis resulted in a single precipitin arc against anti-fluorescein and Anti-Mouse Serum. Reactivity is observed against native Mouse IgG by both Western blot and ELISA.
Labeling Ratio 3.6
Target Name (Antigen)
Application Notes

Immunohistochemistry Dilution: User Optimized

Application Note: Fluorescent Mouse TrueBlot® Antibody Fluorescein may also be used for detection in immunoassays that do not employ immunoprecipitation. Fluorescein Conjugated Antibodies are designed for immunofluorescence microscopy, fluorescence based plate assays (FLISA) and fluorescent western blotting. This product is also suitable for multiplex analysis, including multicolor imaging, utilizing various commercial platforms. Fluorescent Mouse TrueBlot® Antibody Fluorescein 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 2.5mls/blot will yield enough reagent for 40 blots. Note that there are three key procedural considerations: 1. Protein A or G beads may be used with the mouse, goat and sheep TrueBlot secondaries, but not with the rabbit TrueBlot secondary. Use of protein A or G beads with the rabbit TrueBlot will result in contaminating bands. 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. Note: To achieve best results when detecting mouse IgG1 subtypes, we recommend performing a dot blot or titration to determine the optimal dilution factor for your desired application. 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: User Optimized

Western Blot Dilution: 1:1000

IF Microscopy Dilution: 1:200

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-Mouse Ig Fluorescein (ABIN6698838) Immunofluorescence of α-tubulin using FITC-conjugated Fluorescent anti-mouse IgG Immu...
Image no. 2 for Fluorescent TrueBlot®: Anti-Mouse Ig Fluorescein (ABIN6698838) Fluorescein: Anti-Mouse Ig Fluorescein Conjugated Western Blot of Fluorescent: Anti-M...
Image no. 3 for Fluorescent TrueBlot®: Anti-Mouse Ig Fluorescein (ABIN6698838) Mouse IP / Western Blot: Caspase 7 was immunoprecipitated from 0.5 ml of 1x10e7 Jurka...