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Kits  >  Chemiluminescent Western Blot Kits
 

Mouse TrueBlot® Western Blot Kit

Rat Monoclonal eB144
Code

88-8887-31
Size

1 Kit
Price

$249.00
Availability

Ships next business day.
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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.
Shipping & Handling Fees Apply.

Shipping:
$85.00 to United States
Handling:
$20.00
Synonyms: HRP, TrueBlot, HRP TrueBlot ULTRA, Peroxidase TrueBlot, TrueBlot for IP/WB, TrueBlot for immunoprecipitation, TrueBlot for western blotting

Mouse TrueBlot® Western Blot Kit Properties

Description
Mouse TrueBlot® Western Blot Kit - 88-8887-31
Host
Rat
Target Species
Mouse
Known Cross Reactivity
Mouse
Clonality
Monoclonal eB144
Application
Western Blot : 1:1000
ImmunoPrecipitation: Use with TrueBlot® Anti-Mouse Ig IP Beads Cat #( 00-8811-25)
Product Type
Kit
Kit Type
Immunoprecipitation Kit
Physical State
Liquid (sterile filtered)
Shipping condition
Wet Ice
Format
IgG
Buffer
0.01 M Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Stabilizer
0.1 mg/ml Bovine Serum Albumin (BSA) - IgG and Protease free, 50% (v/v) Glycerol
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Mouse TrueBlot® Western Blot Kit Description

Background
The Mouse TrueBlot® Western Blot Kit contains the critical supporting reagents, buffers, and substrates for immunoprecipitation and Western blotting of samples using TrueBlot second step immunoblotting reagents in conjunction with your own primary IP antibody and primary (Mouse IgG) Western blotting antibody. TrueBlot technology enables unhindered detection of protein bands of interest which would otherwise be obscured by the presence of reduced and denatured heavy and light chain immunoglobulin in the blot (as detected by the conventional immunoblotting HRP anti-mouse IgG reagent). Mouse IgG TrueBlot® ULTRA is the unique horseradish peroxidase conjugated anti-mouse IgG immunoblotting second step reagent which enables detection of immunoblotted target protein bands, without hindrance by interfering immunoglobulin heavy and light chains from your IP antibody. Use it in place of your usual HRP anti-mouse IgG immunoblotting second step reagent. It is easy to generate publication-quality IP/WB data with Mouse IgG TrueBlot® ULTRA. Mouse IgG TrueBlot ULTRA 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 ULTRA 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/immunoblotting applications. Applications include studies examining post-translational modification (e.g., phosphorylation or acetylation) or protein-protein interactions.
Storage Condition
Store at 2-8°C except Mouse TrueBlot® ULTRA which should be stored at -20°C. This product is guaranteed for 6 months upon receipt, when handled and stored as instructed.
Application Note
Mouse IgG TrueBlot® ULTRA is provided as 1000X solution. In order 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, cat. 18-8817-31 (50µL) will yield enough reagent for 20 blots. Mouse IgG TrueBlot® ULTRA. An HRP-conjugated second step reagent reacting with mouse IgGs for optimal signal detection in immunoprecipitation/immunoblotting experiments. 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. Milk should be used as the blocking protein for the immunoblot. A sample of Anti-alpha-Tubulin (MOUSE) Monoclonal Antibody Cat #(200-301-880S), has been provided to be used as a loading control at the user’s discretion. Dilutions are to be optimized by the user. The Mouse TrueBlot® Western Blot Kit components are sufficient for 20-25 miniblots. Components: 1. Mouse IgG TrueBlot ULTRA: 50 µl. 18-8817-31 2. TrueBlot Enhancer Solution: 25 ml 3. TrueBlot Blocker: 10 g 4. TrueBlot Assay Buffer: 30 ml. 20X 5. TrueBlot Substrate A: 12.5 ml 6. TrueBlot Substrate B: 12.5 ml 7. Western Blot Incubation Tray 8. Anti-alpha-Tubulin (MOUSE) Monoclonal Antibody - 200-301-880S
Purity/Specifity
Mouse TrueBlot® ULTRA Antibody Peroxidase 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.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 326, Gilbertsville, Pennsylvania, USA.
General Reference
Kong, D., L. Xu, Y. Yu, W. Zhu, D.W. Andrews, Y. Yoon, and T.H. Kuo. 2005. Regulation of Ca2+-induced permeability transition by BCL-2 is antagonized by Drp1 and hFis1. Molecular and Cellular Biochemistry. 272: 187-199. (Rabbit IgG TrueBlot, PubMed) DiPerna, G., J. Stack, A.G. Bowie, A. Boyd, G. Kotwal, Z. Zhang, S. Arvikar, E. Latz, K.A. Fitzgerald, and W.L. Marshall. 2004. Poxvirus protein N1L targets the I-kappaB Kinase complex, inhibits signaling to NF-kappaB by the Tumor Necrosis Factor superfamily of receptors, and inhibits NF-kappaB and IRF3 signaling by Toll-like Receptors. J. Biol. Chem. 279: 36570-36578. (Rabbit IgG TrueBlot, PubMed) Zhang, X., Y. Ozawa, H. Lee, Y. Wen, T. Tan, B. Wadzinski, and E. Seto. 2005. Histone deacetylase 3 (HDAC3) activity is regulated by interaction with protein serine/threonine phosphatase 4. Genes & Development. 19: 827-839. (Rabbit IgG TrueBlot, PubMed) Lehtonen, S., E. Lehtonen, K. Kudlicka, H. Holthöfer, and M.G. Farquhar. 2004. Nephrin Forms a Complex with Adherens Junction Proteins and CASK in Podocytes and in Madin-Darby Canine Kidney Cells Expressing Nephrin. Am J Pathol. 165:923-936. (Rabbit IgG TrueBlot, PubMed) Tyagi A, Agarwal C, Harrison G, Glode LM, Agarwal R. 2004. Silibinin causes cell cycle arrest and apoptosis in human bladder transitional cell carcinoma cells by regulating CDKI-CDK-cyclin cascade, and caspase 3 and PARP cleavages. Carcinogenesis. 25: 1711-20. (Mouse IgG TrueBlot, PubMed)
Specific Reference
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Product Type
Secondary Antibodies;
Applications
WB
Reacts With
Mouse
Catalog Number
18-8817-30

Product Type
Secondary Antibodies;
Applications
WB
Reacts With
Mouse
Catalog Number
18-8817-31

Product Type
Secondary Antibodies;
Applications
WB
Reacts With
Mouse
Catalog Number
18-8817-33

Product Type
Kits;
Applications
WB, IP
Reacts With
Mouse
Catalog Number
00-8811-25

Product Label

 

Description
HRP TrueBlot® ULTRA for IP/WB
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry