Anti-CCT2 antibody [EPR4084]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCT2 antibody [EPR4084] (AB92746)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling CCT2 with purified ab92746 at 1/24000 dilution (0.012 μg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCT2 antibody [EPR4084] (AB92746)

ab92746, unpurified, at a 1/100 dilution, staining CCT2 in formalin fixed, paraffin embedded (1) Human colon tissue and (2) Human kidney tissue by Immunohistochemistry. Detection : DAB staining.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CCT2 antibody [EPR4084] (AB92746)

Immunocytochemistry/Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling CCT2 with Purified ab92746 at 1 : 500 dilution (0.6 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1 : 200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CCT2 antibody [EPR4084] (AB92746)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells, labeling CCT2 with Purified ab92746 at 1/30 dilution (10μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CCT2 antibody [EPR4084] (AB92746)

Overlay histogram showing HeLa cells stained with ab92746, unpurified, (red line), unpurified. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92746, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CCT2 antibody [EPR4084] (AB92746)

ab92746, unpurified, at a 1/100 dilution, staining CCT2 in HeLa cells by Immunofluorescence.

• IP

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Immunoprecipitation - Anti-CCT2 antibody [EPR4084] (AB92746)

Purified ab92746 at 1 : 30 dilution (2μg) immunoprecipitating CCT2 in HeLa whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab92746 + HeLa whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab92746 in HeLa whole cell lysate.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 57 kDa

All lanes:

Immunoprecipitation - Anti-CCT2 antibody [EPR4084] (ab92746)

Predicted band size: 57 kDa

false

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCT2 antibody [EPR4084] (AB92746)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling CCT2 with purified ab92746 at 1/24000 dilution (0.012 μg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCT2 antibody [EPR4084] (AB92746)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling CCT2 with purified ab92746 at 1/24000 dilution (0.012 μg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• WB

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Western blot - Anti-CCT2 antibody [EPR4084] (AB92746)

Blocking/Diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-CCT2 antibody [EPR4084] (ab92746) at 1/10000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

Human brain lysate at 15 µg

Lane 3:

Mouse brain lysate at 15 µg

Lane 4:

Rat brain lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 57 kDa

Observed band size: 57 kDa

false

• WB

Unknown

Western blot - Anti-CCT2 antibody [EPR4084] (AB92746)

All lanes:

Western blot - Anti-CCT2 antibody [EPR4084] (ab92746) at 1/10000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

MCF-7 cell lysate at 10 µg

Lane 3:

Daudi cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 57 kDa

false