Anti-Coronin-1C antibody [EPR25365-24] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Coronin-1C antibody [EPR25365-24] - BSA and Azide free (AB283724)

This data was developed using ab283693, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human lymphoid nodule tissue labelling Coronin-1C with ab283693 at 1/2000 (0.301 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human lymphoid nodule (PMID : 20690162). The section was incubated with ab283693 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Coronin-1C antibody [EPR25365-24] - BSA and Azide free (AB283724)

This data was developed using ab283693, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast tissue labelling Coronin-1C with ab283693 at 1/2000 (0.301 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human breast (PMID : 30065298). The section was incubated with ab283693 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

Lab

Western blot - Anti-Coronin-1C antibody [EPR25365-24] - BSA and Azide free (AB283724)

Blocking buffer and concentration : ½ volume of Odyssey Blocking Buffer (TBS)+ ½ volume of 0.1% TBS                            Diluting buffer and concentration : ½ volume of Odyssey Blocking Buffer (TBS)+ ½ volume of 0.1% TBST    False colour image of Western blot : Anti-Coronin-1C antibody [EPR25365-24] (ab283693) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab283693 was shown to bind specifically to Coronin-1C. A band was observed at 53 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in Coronin-1C knockout cell line ab266381 (knockout cell lysate (ab258377). To generate this image, wild-type and Coronin-1C knockout HEK-293T cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution. This data was developed using ab283693, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Coronin-1C antibody [EPR25365-24] (<a href='/en-us/products/primary-antibodies/coronin-1c-antibody-epr25365-24-ab283693'>ab283693</a>) at 1/1000 dilution

Lane 1:

Wild-type 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CORO1C (Coronin-1C) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-coro1c-coronin-1c-knockout-hek-293t-cell-lysate-ab258377'>ab258377</a>) at 20 µg

Lane 2:

Western blot - Human CORO1C (Coronin-1C) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-coro1c-coronin-1c-knockout-hek-293t-cell-line-ab266381'>ab266381</a>)

Lane 2:

Western blot - Human CORO1C (Coronin-1C) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-coro1c-coronin-1c-knockout-hek-293t-cell-lysate-ab258377'>ab258377</a>)

Secondary

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Coronin-1C antibody [EPR25365-24] - BSA and Azide free (AB283724)

This data was developed using ab283693, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labelling Coronin-1C with ab283693 at 1/2000 (0.301 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human breast cancer (PMID : 30065298). The section was incubated with ab283693 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-Coronin-1C antibody [EPR25365-24] - BSA and Azide free (AB283724)

This data was developed using ab283693, the same antibody clone in a different buffer formulation.

Coronin-1C was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 10 ug with ab283693 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283693 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 10 ug

Lane 2 : ab283693 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283693 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 minutes

All lanes:

Immunoprecipitation - Anti-Coronin-1C antibody [EPR25365-24] (<a href='/en-us/products/primary-antibodies/coronin-1c-antibody-epr25365-24-ab283693'>ab283693</a>)

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

• WB

Lab

Western blot - Anti-Coronin-1C antibody [EPR25365-24] - BSA and Azide free (AB283724)

This data was developed using ab283693, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

Exposure time : 3 minutes

All lanes:

Western blot - Anti-Coronin-1C antibody [EPR25365-24] (<a href='/en-us/products/primary-antibodies/coronin-1c-antibody-epr25365-24-ab283693'>ab283693</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

A549 (human lung carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate at 20 µg

Lane 4:

HL-1 (mouse atrial muscle cell), whole cell lysate at 20 µg

Lane 5:

C6 (rat glial tumor glial cell), whole cell lysate at 20 µg

Lane 6:

Human heart tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

• WB

Lab

Western blot - Anti-Coronin-1C antibody [EPR25365-24] - BSA and Azide free (AB283724)

This data was developed using ab283693, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

This antibody has no cross-reaction with human Coronin-1B, Coronin-6 and Coronin-1A.

All the recombinant proteins were made in-house.

All the samples are E.coli extracts containing Coronin-1C, Coronin-1B, Coronin-6 or Coronin-1A recombinant protein respectively.

Exposure time : 26 seconds

All lanes:

Western blot - Anti-Coronin-1C antibody [EPR25365-24] (<a href='/en-us/products/primary-antibodies/coronin-1c-antibody-epr25365-24-ab283693'>ab283693</a>) at 1/1000 dilution

Lane 1:

His-tagged human Coronin-1C recombinant protein at 0.005 µg

Lane 2:

His-tagged human Coronin-1B recombinant protein at 0.005 µg

Lane 3:

His-tagged human Coronin-6 recombinant protein at 0.005 µg

Lane 4:

His-tagged human Coronin-1A recombinant protein at 0.005 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa

false

• WB

Lab

Western blot - Anti-Coronin-1C antibody [EPR25365-24] - BSA and Azide free (AB283724)

This data was developed using ab283693, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer concentration : 5% NFDM/TBST

Exposure time 26 seconds

All lanes:

Western blot - Anti-Coronin-1C antibody [EPR25365-24] - BSA and Azide free (ab283724) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

Hela transfected with CORO1C siRNA 1 whole cell lysate at 20 µg

Lane 3:

Hela transfected with CORO1C siRNA 2 whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 53 kDa

Observed band size: 53 kDa

false