重组Anti-CEACAM6抗体[EPR23956-80]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CEACAM6 antibody [EPR23956-80] (AB275022)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized BxPC-3 BxPC-3 (human pancreas adenocarcinoma cell) cells labelling CEACAM6 with ab275022 at 1/500 (1.048 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous and cytoplamic staining in BxPC-3 cell line. Low expression control : LNCaP : PMID : 16204051. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM6 antibody [EPR23956-80] (AB275022)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling CEACAM6 with ab275022 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human spleen. The section was incubated with ab275022 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CEACAM6 antibody [EPR23956-80] (AB275022)

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling CEACAM6 with ab275022 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human colon cancer (PMID : 27284373). The section was incubated with ab275022 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 ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

• Flow Cyt

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Flow Cytometry - Anti-CEACAM6 antibody [EPR23956-80] (AB275022)

Flow cytometric analysis of LNCaP (human prostate carcinoma epithelial cell, Left) / BxPC-3 (human pancreas adenocarcinoma epithelial cell, Right) cells labelling CEACAM6 with ab275022 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Negative control : LNCaP.

Gated on viable cells.

• IP

Unknown

Immunoprecipitation - Anti-CEACAM6 antibody [EPR23956-80] (AB275022)

CEACAM6 was immunoprecipitated from 0.35 mg BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate with ab275022 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275022 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab275022 IP in BxPC-3 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab275022 in BxPC-3 whole cell lysate

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-CEACAM6 antibody [EPR23956-80] (ab275022)

Predicted band size: 37 kDa

Observed band size: 40-100 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-CEACAM6 antibody [EPR23956-80] (AB275022)

CEACAM6 was immunoprecipitated from 0.35 mg A549 (human lung carcinoma epithelial cell) whole cell lysate with ab275022 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab275022 at 1/2000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : A549 (human lung carcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab275022 IP in A549 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab275022 in A549 whole cell lysate

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-CEACAM6 antibody [EPR23956-80] (ab275022)

Predicted band size: 37 kDa

Observed band size: 40-100 kDa

false

• WB

Unknown

Western blot - Anti-CEACAM6 antibody [EPR23956-80] (AB275022)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-2 : 15 seconds; Lane3 : 3 seconds; Lane 4 : 26 seconds.

The molecular weight observed is consistent with what has been described in the literature (PMID : 19244123, 28892050, 16204051).

Low expression control : LNCaP (PMID : 16204051).

All lanes:

Western blot - Anti-CEACAM6 antibody [EPR23956-80] (ab275022) at 1/1000 dilution

Lane 1:

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

Lane 2:

LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 37 kDa

Observed band size: 40-100 kDa

false

• WB

Unknown

Western blot - Anti-CEACAM6 antibody [EPR23956-80] (AB275022)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 26 seconds.

The molecular weight observed is consistent with what has been described in the literature (PMID : 19244123, 28892050, 16204051).

All lanes:

Western blot - Anti-CEACAM6 antibody [EPR23956-80] (ab275022) at 1/1000 dilution

All lanes:

Human lung tissue lysate at 20 µg

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 37 kDa

Observed band size: 40-100 kDa

false

• WB

Unknown

Western blot - Anti-CEACAM6 antibody [EPR23956-80] (AB275022)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 15 seconds.

The other family members have lower homology than the tested proteins.

All lanes:

Western blot - Anti-CEACAM6 antibody [EPR23956-80] (ab275022) at 1/1000 dilution

Lane 1:

His-tagged human CEACAM6 recombinant protein, 20ng

Lane 2:

His-tagged human CEACAM1 recombinant protein (aa35-428), 20ng

Lane 3:

His-tagged human CEACAM5 recombinant protein (aa35-685), 20ng

Secondary

All lanes:

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

Predicted band size: 37 kDa

false