Antibodies to alpha and beta Crystallins - FocusOn 038

Introduction: The α-crystallin superfamily and βγ-crystallins compose the major superfamily of proteins present in the crystalline lens. Beta- and gamma-crystallins are similar in sequence, structure and domains topology, and thus have been grouped together as a protein superfamily called βγ-Crystallins.

Alpha-crystallins, composed of alpha A (~19 kDa) and alpha B (~19.2 kDa) subunits, are major water-soluble proteins accounting for almost 50% of total protein in the mammalian transparent eye lens and they are also found in a variety of other tissues (1). Alpha-crystallins are also referred to as small heat shock proteins, since they are induced by increased temperature in a variety of organisms (2). The alpha-crystallins have sequence homology as well as structural and functional similarities with the small Hsp´s such as Hsp25/27 (3). Most alpha-crystallins have four common structural and functional features: (1) molecular weight between 12 and 43kDa; (2) the formation of large oligomeric complexes composed of alpha A-crystallin, alpha B-crystallin and Hsp25/27; (3) the moderately conserved alpha-crystallin domain in the central region of the protein; and (4) molecular chaperone activity (2,4). The alpha-crystallin domain, comprised of approximately 90 residues, is bound by variable N-terminal and C-terminal extensions and is involved in oligomer assembly. Oligomers can reach 800kDa or more and are dynamic, exhibiting subunit exchanges and organizational plasticity, possibly leading to functional diversity. Phosphorylation of serine residues occurs during development and in response to stress, and usually decreases oligomer size (4). Chaperone activity requires, and is modulated by oligomerization and is limited to binding unfolded intermediates to prevent irreversible aggregation (2,4). Other proposed functions include a role in membrane stabilization (2) and modulation of intermediate filament organization during physiological stress and neurodegenerative disease (5).

Antibody Tools for the Detection of Crystallins

Acris Antibodies offers an antibody panel to both alpha-crystallin isoforms and to beta-crystallin; antibodies to the phosphorylated forms of alpha B-crystallin are also available. These monoclonal or polyclonal antibodies can be used in different immunological detection methods like ELISA, Western blotting or immunohistochemistry.



Western blotting of Crystallin alpha A in bovine lens extract using SP5097P

Fig.1 – 4: Western blotting of Crystallin alpha A in bovine lens extract using SP5097P in Fig. 1 and phospho- (lane 1) versus non-phospho recombinant Crystallin alpha B (lane 2) in Fig. 2-4 using SP5098P (Fig. 2), SP5099P (Fig. 3) and SP5100P (Fig. 4).

 

Western blot analysis using antibody SM6011

Fig. 5: Western blot analysis using antibody SM6011
Recombinant alpha-crystallin A (aA) and the extract of mouse eye (ME) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with anti-human alpha-crystallin B (1:1,000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.

 

 

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Literature

  1. Augusteyn, R. C., et al. (1992) Exp. Eye Res. 54:219-228
  2. Narberhaus, F. (2002) Microbiol. Mol. Biol. Rev. 66(1): 64-93.
  3. Merck, K. B.,et al. (1993) J. Biol. Chem. 268: 1046-1052.
  4. MacRae, T.H. (2000) Cell Mol. Life Sci. 57(6): 899-913.
  5. Head, M.W. and Goldman, J.E. (2000) Neuropathol. Appl. Neurobiol. 26(4): 304-312.
  6. van Noort, J.M., et al., 1995, Nature, 375, 798-801
  7. Fujii N., et al., (2003) Mol. Vis. 9: 915-22
  8. Laksanalamai P., et al., (2004) Extremophiles 8: 1-11
  9. De S, Rabin DM, Salero E, Lederman PL, Temple S, Stern JH. (2007) Arch. Ophthalmol. 125(5): 641-5.
  10. Ghosh JG, Shenoy AK, Clark JI. (2007) Biochemistry 46(21): 6308-17

Available Antibody Panel to Crystallin
Anti alpha Crystallin, Crystallin alpha A antibody; Crystallin alpha B antibody, anti beta Crystallin; anti gamma Crystallin; anti zeta Crystallin



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Primary Antibodies

Catalog No. Host Iso. Clone Pres. React. Applications  

Alpha-crystallin A chain / CRYA1 antibody

Western blot analysis of AlphaA Crystallin in bovine tissues at a 1:1000 dilution of the antibody Mouse IgG1 1H3.B8 Aff - Purified Bov, Hu, Ms, Rt E, WB
50 µg / €220.00
  Acris Antibodies GmbH

Alpha-crystallin A chain / CRYA1 antibody

Western blot analysis: Mouse eye extracts and recombinant proteins (Crystallin-alpha A and B) were resolved by electrophoresis, transferred to PVDF membrane and probed with anti-Crystallin alpha A (1:1000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a DAP detection system. This antibody does not show cross-activity about Crystallin alpha B.            Lane A: Crystallin-alpha A recombinant protein, Lane B: Crystallin-alpha B recombinant protein, Lane C: Mouse eye lysates Mouse IgG1 c9F2 Purified Hu, Ms, Rt E, WB
0.1 ml / €290.00
  Acris Antibodies GmbH

Alpha-crystallin A chain / CRYA1 antibody

Western blot analysis: Mouse eye extracts and recombinant proteins (Crystallin-alpha A and B) were resolved by electrophoresis, transferred to PVDF membrane and probed with anti-Crystallin alpha A (1:1000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a DAP detection system. This antibody does not show cross-activity about Crystallin alpha B.            Lane A: Crystallin-alpha A recombinant protein, Lane B: Crystallin-alpha B recombinant protein, Lane C: Mouse eye lysates Mouse IgG1 c9F2 Purified Hu, Ms, Rt E, WB
50 µl / €230.00
  Acris Antibodies GmbH

Alpha-crystallin B chain / CRYA2 antibody

NULL Mouse IgG1 3A10.C9 Aff - Purified Bov, Hu E, P, WB
50 µg / €220.00
  Acris Antibodies GmbH

Alpha-crystallin B chain / CRYA2 antibody

Western blot analysis of AlphaB Crystallin in bovine cell lines using a 1:1000 dilution of the antibody Mouse IgG1 1A7.D5 Aff - Purified Bov, Hu E, P, WB
50 µg / €220.00
  Acris Antibodies GmbH

Alpha-crystallin B chain / CRYA2 antibody

Alpha-crystallin B chain / CRYA2 Rabbit IgG Serum Bov, Chk, Hu, Ms, Rt C, ICC/IF, WB
50 µl / €210.00
  Acris Antibodies GmbH

Alpha-crystallin B chain / CRYA2 antibody

Western blot analysis: Recombinant α-crystallin A (αA) and the extract of mouse eye (ME) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with anti-human α-crystallin B antibody (1:1,000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system. Mouse IgG2b 2E8 Purified Hu, Ms E, F, WB
0.1 ml / €310.00
  Acris Antibodies GmbH

Alpha-crystallin B chain / CRYA2 antibody

Western blot analysis: Recombinant α-crystallin A (αA) and the extract of mouse eye (ME) were resolved by SDS-PAGE, transferred to PVDF membrane and probed with anti-human α-crystallin B antibody (1:1,000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system. Mouse IgG2b 2E8 Purified Hu, Ms E, F, WB
50 µl / €230.00
  Acris Antibodies GmbH

Beta-crystallin S (19-37) antibody

Western blot of AP09926PU-N (1:750). Approx. MW of Beta Crystallin is 21 kDa. Rabbit Aff - Purified Hu, Ms, Rt E, IP, WB
0.1 mg / €420.00
  Acris Antibodies GmbH

Beta-crystallin S (C-term) antibody

Beta-crystallin S Rabbit Aff - Purified Hu, Ms, Rt C, E, IP, WB
0.1 mg / €420.00
  Acris Antibodies GmbH

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