Antibodies to CCT - FocusOn 086

Introduction

Chaperones are proteins whose function is to assist other proteins in achieving proper folding. Many chaperones are heat shock proteins, that is, proteins expressed in response to elevated temperatures.

The reason for this behaviour is that protein folding is severely affected by heat, and therefore chaperones act to counteract the potential damage. Protein folding is an essential step in the expression of genetic information, and defects in this process lead to a variety of disease states.

There are several functionally and structurally distinct chaperone systems in the eukaryotic cytosol. The cytosolic chaperonin-containing t-complex polypeptide 1 (CCT) is a molecular chaperone that plays an important role in the folding of proteins in the eukaryotic cytosol. Actin, tubulin, and several other proteins are known to be folded by CCT, and an estimated 15% of newly translated proteins in mammalian cells are folded with the assistance of CCT. CCT differs from other chaperonin family proteins in its subunit composition, which consists of eight subunit species comprising the CCT 16-mer double-ring-like complex. CCT preferentially recognizes quasinative (or partially folded) intermediates, whereas its Escherichia coli homologue GroEL recognizes more unfolded intermediates, especially those displaying hydrophobic surfaces. Molecular evolutionary analyses have suggested that each subunit species has a specific function in addition to contributing to a common ATPase activity. Consistent with this view, it has been suggested that each subunit recognizes specific substrate proteins (or their parts) and that they collectively modulate the ATPase activity of the complex. The overall expression of CCT in mammalian cells is primarily dependent on cell growth, but each subunit exhibits an individual patterns of expression.

Antibody Panel to CCT

Acris Antibodies offers a range of monoclonal antibodies to different polypeptides of the CCT chaperonin molecule complex. These antibodies can be used in Western blot and / or Immunoprecipitation applications.

CCT1

Fig. 1: TCP1 antibody Cat.-No. AP16211PU-N staining of paraffin embedded human testis at 10 µg/ml. Microwaved antigen retrieval with Tris/EDTA buffer pH9, HRP-staining.

Fig. 2: Western blot analysis of HeLa whole cell lysate (lane 1) and HeLa heat stressed whole cell lysate (lane 2) probed with rat anti mouse CCT alpha Cat.-No. SM2173P followed by F(ab')2 rabbit anti Rat IgG:HRP (Cat.-No. SP1021HRP)

CCT2

Fig. 3: Western blot analysis of HeLa whole cell lysate (Lane 1) and HeLa heat stressed whole cell lysate (Lane 2) probed with Rat Anti Human CCT Beta antibody (SM2016P) followed by F(ab')2 Rabbit Anti Rat IgG:HRP (SP1021HRP).

Fig. 4: Immunofluorescence analysis of 3T3-L1 cells using CCT2 mouse mAb (green). Blue: DRAQ5 fluorescent DNA dye. Red: Actin filaments have been labeled with Alexa Fluor-555 phalloidin.

CCT3

Fig. 5: Western blot analysis of CCT3 Antibody (C-term) (Cat.-No. AP17199PU-N) in K562 cell line lysates (35 µg/lane). CCT3 (arrow) was detected using the purified Pab.

Fig. 6: Immunohistochemistry of paraffin-embedded Colon cancer using 10571-1-AP (CCT3 Antibody) at Dilution 1:50 (under 10x lens)

CCT5

Fig. 7: Human Brain, Cortex: Formalin-Fixed, Paraffin-Embedded (FFPE) (Cat.-No. AM31096PU-N)

Fig. 8: Immunofluorescence of monoclonal antibody to CCT5 on HeLa cell. [antibody concentration 10 µg/ml] (cat.-No. AM31096PU-N)

CCT6a

Fig. 9: Hela cell were subjected to SDS PAGE followed by western blot with 19793-1-AP(CCT6A-Specific Antibody) at dilution of 1:600 (Cat.-No. 19793-1-AP)

CCT6b

Fig. 10: CCT6B monoclonal antibody (M01), clone 1A4 Western Blot analysis of CCT6B expression in HL-60 (Cat.-No. L014V1).

CCT7

Fig. 11: Western blot analysis of HeLa whole cell lysate (lane 1) and HeLa heat stressed whole cell lysate (lane 2) probed with rat anti CCT Eta Cat.-No. SM2019P followed by F(ab')2 rabbit anti rat IgG:HRP (Cat.No. SP1021HRP).

Fig. 12: Hela cell were subjected to SDS PAGE followed by western blot with 15994-1-AP (CCT7 Antibody) at dilution of 1:1500

CCT8

Fig. 13: Western blot analysis of HeLa whole cell lysate (lane 1) and HeLa heat stressed whole cell lysate (lane 2) probed with rat anti CCT theta Cat.-No. SM2017P followed by F(ab')2 rabbit anti rat IgG:HRP (Cat.-No. SP1021HRP).

Fig. 14: Immunofluorescent analysis of MCF-7 cells, using CCT8 antibody 12263-1-AP  at 1:25 dilution and Rhodamine-labeled goat anti-rabbit IgG (red).

Literature

Horwich AL, Fenton WA, Chapman E, Farr GW Two families of chaperonin: physiology and mechanism. Annu Rev Cell Dev Biol. 2007;23:115-45.

Liu X, Lin CY, Lei M, Yan S, Zhou T, Erikson RL. CCT chaperonin complex is required for the biogenesis of functional Plk1. Mol Cell Biol. 2005 Jun 25(12):4993-5010.

Melville MW, McClellan AJ, Meyer AS, Darveau A, Frydman J. The Hsp70 and TRiC/CCT Chaperone Systems Cooperate In Vivo To Assemble the Von Hippel-Lindau Tumor Suppressor Complex. Molecular and Cellular Biology 2003 May 23(9): 3141-3151.

Kubota H. Function and regulation of cytosolic molecular chaperone CCT. Vitam Horm. 2002;65(3):13-31.

Llorca O, Martin-Benito J, Grantham J, Ritco-Vonsovici M, Willison KR, Carrascosa JL, Valpuesta JM. The 'sequential allosteric ring' mechanism in the eukaryotic chaperonin-assisted folding of actin and tubulin. EMBO J. 2001 Aug 1;20(15):4065-75.