Anti-Cathelicidin/CLP antibody [RM1363]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Cathelicidin/CLP with ab325356 at 1/2000 (0.246 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen.

The primary antibody was incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissue labeling Cathelicidin/CLP with ab325356 at 1/2000 (0.246 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on granulocytes of human cervical cancer.

The primary antibody was incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Human neutrophils cells labelling Cathelicidin/CLP with ab325356 at 1/500 dilution (0.1ug) / Red compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

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

• IP

Lab

Immunoprecipitation - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Cathelicidin/CLP was immunoprecipitated from 0.35 mg Human spleen tissue lysate with ab325356 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325356 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Cathelicidin/CLP antibody [RM1363] (ab325356) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 10 µg

Lane 2:

ab325356 at 1/30 IP in Human spleen tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325356 in human spleen tissue 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

Observed band size: 19 kDa

false

Exposure time: 10s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Cathelicidin/CLP with ab325356 at 1/2000 (0.246 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen.

The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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)

Lab

Flow Cytometry (Intracellular) - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse bone marrow cells labelling Cathelicidin/CLP with ab325356 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling Cathelicidin/CLP with ab325356 at 1/2000 (0.246 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung.

The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Immunohistochemical analysis of paraffin-embedded Mouse pancreatic cancer tissue labeling Cathelicidin/CLP with ab325356 at 1/2000 (0.246 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on granulocytes of mouse pancreatic cancer.

The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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

Lab

Immunoprecipitation - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Cathelicidin/CLP was immunoprecipitated from 0.35 mg Mouse spleen tissue lysate with ab325356 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325356 at 1/1000 dilution.

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

All lanes:

Immunoprecipitation - Anti-Cathelicidin/CLP antibody [RM1363] (ab325356) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 10 µg

Lane 2:

ab325356 at 1/30 IP in Mouse spleen tissue lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab325356 in mouse spleen tissue 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

Observed band size: 19 kDa

false

Exposure time: 24s

• WB

Lab

Western blot - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

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

Low expression : cerebellum (PMID : 9148921).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Cathelicidin/CLP antibody [RM1363] (ab325356) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Lane 3:

Human small intestine tissue lysate at 20 µg

Lane 4:

Human cerebellum tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/5000 dilution

Observed band size: 19 kDa,36 kDa

false

Exposure time: 48s

• WB

Lab

Western blot - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

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

Low expression : brain (PMID : 9148921).

The identity of the higher MW band at approximately 50 kDa (in lane 2/3 are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lane 1 : 4 seconds; Lane 2 and 3 : 70 seconds

All lanes:

Western blot - Anti-Cathelicidin/CLP antibody [RM1363] (ab325356) at 1/1000 dilution

Lane 1:

Mouse plasma lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate at 20 µg

Lane 3:

Mouse brain tissue 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: 19 kDa,36 kDa

false

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cells labelling Cathelicidin/CLP with ab325356 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

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

Negative control : Mouse primary neuron.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Cathelicidin/CLP with ab325356 at 1/2000 (0.246 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on mouse cerebrum.

The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

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)

Lab

Flow Cytometry (Intracellular) - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling Cathelicidin/CLP with ab325356 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

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

Negative control : Human PBMC.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathelicidin/CLP antibody [RM1363] (AB325356)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Cathelicidin/CLP with ab325356 at 1/2000 (0.246 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on human cerebrum.

The primary antibody was incubated 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins