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TCP1 alpha / CCTA Recombinant Rabbit Monoclonal Antibody [JE48-48]

Usd: 385

Specification

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Catalog# ET7109-79

TCP1 alpha / CCTA Recombinant Rabbit Monoclonal Antibody [JE48-48]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

Conjugation

  • unconjugated

Overview

Product Name

TCP1 alpha / CCTA Recombinant Rabbit Monoclonal Antibody [JE48-48]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human TCP1 alpha / CCTA aa 1-97 / 556.

Species Reactivity

Human, Mouse

Validated Applications

WB, IHC-P, IF-Cell, FC

Target Molecular Weight

Predicted band size: 60 kDa

Positive Control

Daudi cell lysates, NIH/3T3 cell lysate, Mouse testis tissue lysate, human liver cancer tissue, human testis tissue, Jurkat.

Conjugation

unconjugated

Clone Number

JE48-48

RRID

AB_3070954

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:200-1:1,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

The protein encoded by this gene is a molecular chaperone that is a member of the chaperonin containing TCP1 complex (CCT), also known as the TCP1 ring complex (TRiC). This complex consists of two identical stacked rings, each containing eight different proteins. Unfolded polypeptides enter the central cavity of the complex and are folded in an ATP-dependent manner. The complex folds various proteins, including actin and tubulin. Alternate transcriptional splice variants of this gene, encoding different isoforms, have been characterized. In addition, three pseudogenes that appear to be derived from this gene have been found. The TRiC complex mediates the folding of WRAP53/TCAB1, thereby regulating telomere maintenance. As part of the TRiC complex may play a role in the assembly of BBSome, a complex involved in ciliogenesis regulating transports vesicles to the cilia. The TRiC complex plays a role in the folding of actin and tubulin (Probable).

Background References

1. Freund A et al. Proteostatic control of telomerase function through TRiC-mediated folding of TCAB1. Cell 159:1389-1403 (2014).

2. Seo S et al. BBS6, BBS10, and BBS12 form a complex with CCT/TRiC family chaperonins and mediate BBSome assembly. Proc. Natl. Acad. Sci. U.S.A. 107:1488-1493 (2010).

Sequence Similarity

Belongs to the TCP-1 chaperonin family.

Subcellular Location

Cytoplasm. Cytoskeleton.

UNIPROT

SwissProt: P17987 Human

SwissProt: P11983 Mouse

Synonyms

AI528772 antibody

c-cpn antibody

CCT alpha antibody

CCT antibody

CCT-alpha antibody

CCT1 antibody

Ccta antibody

CCTalpha antibody

D6S230E antibody

MGC133746 antibody

Expand

Images

  • Western blot analysis of TCP1 alpha / CCTA on different lysates with Rabbit anti-TCP1 alpha / CCTA antibody (<a href="/products/ET7109-79" style="font-weight: bold;text-decoration: underline;">ET7109-79</a>) at 1/5,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate<br />Lane 2: Mouse testis tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 60 kDa<br />Observed band size: 60 kDa<br /><br />Exposure time: 25 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET7109-79" style="font-weight: bold;text-decoration: underline;">ET7109-79</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of TCP1 alpha / CCTA on different lysates with Rabbit anti-TCP1 alpha / CCTA antibody (ET7109-79) at 1/5,000 dilution.

    Lane 1: NIH/3T3 cell lysate
    Lane 2: Mouse testis tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 60 kDa
    Observed band size: 60 kDa

    Exposure time: 25 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7109-79) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-TCP1 alpha / CCTA antibody (<a href="/products/ET7109-79" style="font-weight: bold;text-decoration: underline;">ET7109-79</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-79" style="font-weight: bold;text-decoration: underline;">ET7109-79</a>) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-TCP1 alpha / CCTA antibody (ET7109-79) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-79) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-TCP1 alpha / CCTA antibody (<a href="/products/ET7109-79" style="font-weight: bold;text-decoration: underline;">ET7109-79</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET7109-79" style="font-weight: bold;text-decoration: underline;">ET7109-79</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-TCP1 alpha / CCTA antibody (ET7109-79) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7109-79) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunocytochemistry analysis of Jurkat cells labeling TCP1 alpha / CCTA with Rabbit anti-TCP1 alpha / CCTA antibody (<a href="/products/ET7109-79" style="font-weight: bold;text-decoration: underline;">ET7109-79</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TCP1 alpha / CCTA antibody (<a href="/products/ET7109-79" style="font-weight: bold;text-decoration: underline;">ET7109-79</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of Jurkat cells labeling TCP1 alpha / CCTA with Rabbit anti-TCP1 alpha / CCTA antibody (ET7109-79) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-TCP1 alpha / CCTA antibody (ET7109-79) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of Jurkat cells labeling TCP1 alpha / CCTA.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET7109-79" style="font-weight: bold;text-decoration: underline;">ET7109-79</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Jurkat cells labeling TCP1 alpha / CCTA.

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET7109-79, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Western blot analysis of TCP1 alpha / CCTA on different lysates with Rabbit anti-TCP1 alpha / CCTA antibody (<a href="/products/ET7109-79" style="font-weight: bold;text-decoration: underline;">ET7109-79</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate<br />Lane 2: K-562 (Human chronic myelogenous leukemia cell) cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br />Exposure time: 10 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/ET7109-79" style="font-weight: bold;text-decoration: underline;">ET7109-79</a>, 1/5,000 in 5% NFDM/TBST, overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 60 kDa<br />Observed band size: 60 kDa

    Western blot analysis of TCP1 alpha / CCTA on different lysates with Rabbit anti-TCP1 alpha / CCTA antibody (ET7109-79) at 1/5,000 dilution.

    Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
    Lane 2: K-562 (Human chronic myelogenous leukemia cell) cell lysate

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 10 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: ET7109-79, 1/5,000 in 5% NFDM/TBST, overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 60 kDa
    Observed band size: 60 kDa

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