Anti-DMBT1 antibody [EPR25722-10] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DMBT1 antibody [EPR25722-10] - BSA and Azide free (AB316186)

This data was developed using ab316185, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human ileum tissue labeling DMBT1 with ab316185 at 1/2000 (0.242 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Cytoplasmic staining of the crypts and very basal parts of human ileum (PMID : 32436776).
The section was incubated with ab316185 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-DMBT1 antibody [EPR25722-10] - BSA and Azide free (AB316186)

This data was developed using ab316185, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell, Left) / Capan-1 (human pancreatic adenocarcinoma epithelial cell, Right) cells labelling DMBT1 with ab316185 at 1/50 dilution (1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : MCF7.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DMBT1 antibody [EPR25722-10] - BSA and Azide free (AB316186)

This data was developed using ab316185, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human duodenum tissue labeling DMBT1 with ab316185 at 1/2000 (0.242 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Cytoplasmic staining of the crypts and very basal parts of human duodenum(PMID : 32436776).
The section was incubated with ab316185 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 was performed 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-DMBT1 antibody [EPR25722-10] - BSA and Azide free (AB316186)

This data was developed using ab316185, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling DMBT1 with ab316185 at 1/2000 (0.242 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on human liver (PMID : 32436776).
The section was incubated with ab316185 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-DMBT1 antibody [EPR25722-10] - BSA and Azide free (AB316186)

This data was developed using ab316185, the same antibody clone in a different buffer formulation.

DMBT1 was immunoprecipitated from 0.35 mg Capan-1 (human pancreas adenocarcinoma epithelial cell) whole cell lysate with ab316185 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316185 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-DMBT1 antibody [EPR25722-10] (<a href='/en-us/products/primary-antibodies/dmbt1-antibody-epr25722-10-ab316185'>ab316185</a>) at 1/1000 dilution

Lane 1:

Capan-1 (human pancreas adenocarcinoma epithelial cell) whole cell lysate

Lane 2:

<a href='/en-us/products/primary-antibodies/dmbt1-antibody-epr25722-10-ab316185'>ab316185</a> at 1/30 IP in Capan-1 (human pancreas adenocarcinoma epithelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/dmbt1-antibody-epr25722-10-ab316185'>ab316185</a> in Capan-1 whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 41s

• WB

Supplier Data

Western blot - Anti-DMBT1 antibody [EPR25722-10] - BSA and Azide free (AB316186)

This data was developed using ab316185, the same antibody clone in a different buffer formulation.

Nagative control : MCF7(PMID : 32436776)

The molecular weight observed is consistent with what has been described in the literature (PMID : 12015815).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-DMBT1 antibody [EPR25722-10] (<a href='/en-us/products/primary-antibodies/dmbt1-antibody-epr25722-10-ab316185'>ab316185</a>) at 1/1000 dilution

Lane 1:

Capan-1 (human pancreatic adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 270 kDa,330 kDa,36 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-DMBT1 antibody [EPR25722-10] - BSA and Azide free (AB316186)

This data was developed using ab316185, the same antibody clone in a different buffer formulation.

Low expression : U-937, THP-1.

The molecular weight observed is consistent with what has been described in the literature (PMID : 12015815).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-DMBT1 antibody [EPR25722-10] (<a href='/en-us/products/primary-antibodies/dmbt1-antibody-epr25722-10-ab316185'>ab316185</a>) at 1/1000 dilution

Lane 1:

SUM149PT (human breast adenocarcinoma cell) whole cell lysate at 20 µg

Lane 2:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Lane 3:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 270 kDa,330 kDa,36 kDa

false

Exposure time: 70s

• WB

Supplier Data

Western blot - Anti-DMBT1 antibody [EPR25722-10] - BSA and Azide free (AB316186)

This data was developed using ab316185, the same antibody clone in a different buffer formulation.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-DMBT1 antibody [EPR25722-10] (<a href='/en-us/products/primary-antibodies/dmbt1-antibody-epr25722-10-ab316185'>ab316185</a>) at 1/1000 dilution

All lanes:

His-tagged human DMBT1 recombinant protein fragment at 10 ng

Secondary

All lanes:

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

Observed band size: 60 kDa

false

Exposure time: 10s