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Monoclonal Antibodies

Anti-MBP [BBMBP23.42] monoclonal antibody

Invented by Brian Burke
Invented at A*STAR Accelerate Technologies Pte Ltd
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Image thumbnail for Anti-MBP [BBMBP23.42] monoclonal antibody
Image thumbnail for Anti-MBP [BBMBP23.42] monoclonal antibody
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Shown are confocal images of Hela cells stably expressing green fluorescence protein (GFP)-tagged MBP (amino acids residues 27-392) that were fixed in 4% PFA (upper panel) or MeOH (lower panel) and stained for MBP (in red) with anti-MBP BBMBP23.42 monoclonal antibody. GFP is shown in green. The nuclei were counter-stained with Hoechst.
Western blot analysis of bacterially expressed MBP with BBMBP23.42 antibody. E.coli BL21 transformed with constructs indicated above were mock (-) or IPTG (+) induced for short time. After induction the whole-cell lysates were separated on SDS-PAGE gel, transferred to nitrocellulose membrane and blotted with BBMBP23.42 antibody. Both primary and secondary antibodies were diluted in blocking buffer (5% nonfat dry milk resuspended in 0.1% Tween and 1X PBS). The predicted molecular weights of constructs are: GST-cyren (112AA), 38 kDA; MBP, 45 kDA; MBP-ZMPSTE24 (113AA), 57 kDa; MBP-Sun1 (64AA), 48 kDa. The length of the fragment fused to a tag is indicated by number of amino acids residues (AA).
BBMBP23.42 detecting MBP expressed in a mammalian cell line by Western blot. Whole cell lysate from Hela cell line stably expressing GFP-tagged GST (Hela GFP-GST) or MBP (Hela GFP-MBP) were separated on SDS-PAGE gel, transferred to nitrocellulose membrane and blotted with BBMBP23.42 antibody. Both primary and secondary antibodies were diluted in blocking buffer (5% nonfat dry milk resuspended in 0.1% Tween and 1X PBS). The predicted molecular weight of GFP-GST and MBP is 53 kDa and 67 kDa respectively. The same blot was stripped and probed for a loading control (beta actin, lower panel).
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Catalogue Number 154470
Applications IF WB
Antigen/Gene or Protein Targets Maltose Binding Protein
Reactivity E.coli
Relevance The E. coli maltose binding protein (MBP) is a product of the malE gene and is part of two multicomponent systems that uptake and sense maltose and higher-order glucose polymers (e.g. maltodextrin). MBP acts in the periplasmic space where it binds with high affinity to maltose/maltodextrin which, in turn binds to (i) a transport apparatus that translocates the MBP ligand across the inner membrane to the cytoplasm and (ii) chemoreceptors causing activation of a signaling pathway that directs taxis toward maltose (1).

MBP is widely used as an affinity tag for purification of recombinant proteins. The reversible and high-affinity binding of MBP to amylose resins enables recovery of recombinant proteins at high purity in a single purification step. The purification and elution are performed under mild conditions, preserving the activity of the recombinant protein. In addition, MBP might increase expression, proper folding, and solubility, of its fusion partner (2–4).
Host Mouse
Immunogen E.coli maltose binding protein (MBP) fusion construct
Immunogen UniProt ID P0AEX9
Positive Control MBP expressing construct in bacteria or mammalian cells
Subclass IgG2a kappa
Molecular Weight (kDa) MBP precursor polypeptide 43 kDa, mature MBP 42 kDa
Notes Immunocytochemistry/Immunofluorescence: formaldehyde, methanol fixation
Western blot: MBP expressed in mammalian and bacteria systems
Research Area Gene Expression, Genetic Studies Tools

References

There are 4 reference entries for this reagent.

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References: 4 entries

Kimple et al. 2013. Curr Protoc Protein Sci. 73:Unit 9.9. PMID: 24510596.

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Riggs et al. 2001. Curr Protoc Mol Biol. Chapter 16:Unit16.6. PMID: 18265133.

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Kapust et al. 1999. Protein Sci. 8(8):1668-74. PMID: 10452611.

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Boos et al. 1998. Microbiol Mol Biol Rev. 62(1):204-29. PMID: 9529892.

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References: 4 entries

Kimple et al. 2013. Curr Protoc Protein Sci. 73:Unit 9.9. PMID: 24510596.

View  

Riggs et al. 2001. Curr Protoc Mol Biol. Chapter 16:Unit16.6. PMID: 18265133.

View  

Kapust et al. 1999. Protein Sci. 8(8):1668-74. PMID: 10452611.

View  

Boos et al. 1998. Microbiol Mol Biol Rev. 62(1):204-29. PMID: 9529892.

View  

Add a reference

Inventor Information