Maltose Binding Protein (MBP) antibody

Antigen: Maltose Binding Protein (MBP)

Clonality: Polyclonal

Host: Rabbit

Application: Immunohistochemistry (IHC), ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN103913

Additional Information

Immunogen: This antibody was purified from whole rabbit serum prepared by repeated immunizations with the MBP epitope tag recombinant protein.

Format: Lyophilized

Description: IgG Fraction of anti-MALTOSE BINDING PROTEIN (MBP) EPITOPE TAG (Rabbit). Epitope tags are short peptide sequences that are easily recognized by tag-specific antibodies. Due to their small size, epitope tags do not affect the tagged protein's biochemical properties. Most often sequences encoding the epitope tag are included with target DNA at the time of cloning to produce fusion proteins containing the epitope tag sequence. This allows anti-epitope tag antibodies to serve as universal detection reagents for any tag containing protein produced by recombinant means. This means that anti-epitope tag antibodies are a useful alternative to generating specific antibodies to identify, immunoprecipitate or immunoaffinity purify a recombinant protein. The anti-epitope tag antibody is usually functional in a variety of antibody-dependent experimental procedures. Expression vectors producing epitope tag fusion proteins are available for a variety of host expression systems including bacteria, yeast, insect and mammalian cells. Antibodies-Online Immunochemicals produces anti-epitope tag antibodies against many common epitope tags including Myc, GST, GFP, 6X His, MBP, FLAG and HA. Antibodies-Online Immunochemicals also produces antibodies to other tags including FITC, Rhodamine (TRITC), DNP and biotin.

Specificity: This polyclonal anti-MBP tag antibody detects over-expressed proteins containing the MBP epitope tag. To date this antibody has reacted with all MBP tagged proteins so far tested. In western blotting of bacterial extracts the antibody does not cross-react with endogenous proteins.

Application Details

Application Notes: Recommended Dilutions: This product was assayed by immunoblot and was found to be reactive at 1:3,500. For ELISA a dilution of 1:4,000 to 1:16,000 is suggested for detection of MBP containing recombinant proteins or for screening in microtiter plates of recombinants. Use antibodies-online's HRP Anti- RABBIT IgG (H&L) (DONKEY) MX Bv Ch Gt GP Hs Hu Ms Rt & Sh (ABIN101955) as a secondary antibody for detection. The researcher should determine optimal titers for other applications. Figure. Anti-MBP epitope tag polyclonal antibody detects MBP-tagged recombinant proteins by western blot. Polyclonal rabbit-anti-MBP epitope tag at 0.5-1.0 ug/ml was used to detect 1.0 ug of recombinant protein containing the MBP epitope tag. A 4-20% gradient gel was used to separate the protein by SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking the membrane was probed with the primary antibody for 1 h at room temperature followed by washes and reaction with a 1:2500 dilution of IRDye 800 conjugated Gt-a-Rabbit IgG [H&L] (code ABIN102013) for 30 min at room temperature. LICOR's Odyssey Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results. Note: Anti-MBP is optimally suited for monitoring the expression of MBP tagged fusion proteins. As such, anti- MBP/MBP can be used to identify fusion proteins containing the MBP epitope. The antibody recognizes the MBP epitope tag fused to the amino- or carboxy- termini of targeted proteins. This antibody has been tested by ELISA and western blotting against MBP containing recombinant proteins. Although not tested, this antibody is likely functional for immunoprecipitation and immunocytochemistry, and other immunodetection techniques. Maltose binding protein is a bacterial protein, which is often used in protein expression studies because it creates a stable fusion product that does not appear to interfere with the bioactivity of the protein of interest. It also allows for its easy purification from bacterial extracts under mild conditions. Anti-MBP is a companion to the pMAL protein expression system and can be used for the detection and purification of MBP-fusion proteins expressed in E. coli. By Western blot, a band is seen at ~ 42 kDa representing MBP.

Concentration: 1.0 mg/ml (by UV absorbance at 280 nm)

Purity: This IgG purified antibody is directed against MBP and is useful in determining its presence in various assays.

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at 4° C prior to restoration. Restore with 1.0 ml of deionized water (or equivalent). 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. Expiration date is one (1) year from date of restoration.

Research Area: Complement Factors, Receptors, Serum/Plasma Proteins, Inflammation

Restrictions: For Research Use only

Images

Anti-MBP epitope tag polyclonal antibody detects MBP-tagged recombinant proteins by western blot. Polyclonal rabbit-anti-MBP epitope tag at 0.5-1.0 ug/ml was used to detect 1.0 ug of recombinant protein containing the MBP epitope tag. A 4-20% gradient gel was used to separate the protein by SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking the membrane was probed with the primary antibody for 1 h at room temperature followed by washes and reaction with a 1:2500 dilution of IRDye

Simplified diagram of MBP-fusion protein construct using pMal expression vector system.