Anti-CXCL16 antibody [EPR29714-545]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCL16 antibody [EPR29714-545] (AB324591)

Immunohistochemical analysis of paraffin-embedded (A)HEK-293T (human embryonic kidney epithelial cell) transfected with a mouse CXCL16 expression vector containing a his tag and (B)HEK-293T transfected with empty vector containing a his tag labeling CXCL16 with ab324591 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T transfected with a mouse CXCL16 expression vector containing a his tag. No staining on (B) HEK-293T transfected with empty vector containing a his tag.
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 Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCL16 antibody [EPR29714-545] (AB324591)

Immunohistochemical analysis of paraffin-embedded A RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cell pellets treated with 100ng/ml Lipopolysaccharide (LPS) for 24 hours, then 300ng/ml Brefeldin A was added for additional 20 hours and B Untreated RAW 264.7 cell pellets tissue labeling CXCL16 with ab324591 at 1/50 (10.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) RAW 264.7 cell pellets treated with 100ng/ml Lipopolysaccharide (LPS) for 24 hours, then 300ng/ml Brefeldin A was added for additional 20 hours, no staining on (B) Untreated RAW 264.7 cell pellets.
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 Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCL16 antibody [EPR29714-545] (AB324591)

Immunohistochemical analysis of paraffin-embedded A mouse lung treated with 1ug/ml Lipopolysaccharide (LPS) for 16 hours, then 1ug/ml Brefeldin A was added for additional 16 hours and B Untreated mouse lung tissue labeling CXCL16 with ab324591 at 1/50 (10.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) mouse lung treated with 1ug/ml Lipopolysaccharide (LPS) for 16 hours, then 1ug/ml Brefeldin A was added for additional 16 hours, no staining on (B) Untreated mouse lung.
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 Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-CXCL16 antibody [EPR29714-545] (AB324591)

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

The expression of CXCL16 is upregulated in response to LPS treatment (PMID : 31199046).

CXCL16 undergoes cleavage and glycosylation modifications. Consequently, different band sizes corresponding to its distinct forms can be detected : the 28-37 kDa precursor form, the 50 kDa mature form, and the 38 kDa soluble form. Please refer to publications such as PMID : 15004171, PMID : 39436058, PMID : 33800554, PMID : 17467666.

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

All lanes:

Western blot - Anti-CXCL16 antibody [EPR29714-545] (ab324591) at 1/1000 dilution

Lane 1:

Untreated RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 50 µg

Lane 2:

RAW 264.7 treated with 100 ng/ml Lipopolysaccharide (LPS) for 3 hours, then 300 ng/ml Brefeldin A was added for additional 3 hours, whole cell lysate at 50 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: 50 kDa,38 kDa,28-37 kDa,15 kDa,36 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-CXCL16 antibody [EPR29714-545] (AB324591)

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

The expression of CXCL16 is upregulated in response to LPS treatment (PMID : 31199046).

CXCL16 undergoes cleavage and glycosylation modifications. Consequently, different band sizes corresponding to its distinct forms can be detected : the 28-37 kDa precursor form, the 50 kDa mature form, and the 38 kDa soluble form. Please refer to publications such as PMID : 15004171, PMID : 39436058, PMID : 33800554, PMID : 17467666.

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

All lanes:

Western blot - Anti-CXCL16 antibody [EPR29714-545] (ab324591) at 1/1000 dilution

Lane 1:

Untreated J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) whole cell lysate at 50 µg

Lane 2:

J774A.1 treated with 100 ng/ml Lipopolysaccharide (LPS) for 18 hours whole cell lysate at 50 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: 50 kDa,38 kDa,28-37 kDa,15 kDa,36 kDa

false

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-CXCL16 antibody [EPR29714-545] (AB324591)

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

The expression of CXCL16 is upregulated in response to LPS treatment (PMID : 31199046).

CXCL16 undergoes cleavage and glycosylation modifications. Consequently, different band sizes corresponding to its distinct forms can be detected : the 28-37 kDa precursor form, the 50 kDa mature form, and the 38 kDa soluble form. Please refer to publications such as PMID : 15004171, PMID : 39436058, PMID : 33800554, PMID : 17467666.

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

All lanes:

Western blot - Anti-CXCL16 antibody [EPR29714-545] (ab324591) at 1/1000 dilution

Lane 1:

Untreated mouse lung tissue lysate at 50 µg

Lane 2:

Mouse lung treated with 1 ug/ml Lipopolysaccharide (LPS) for 20 hours lysate at 50 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: 50 kDa,38 kDa,28-37 kDa,15 kDa,36 kDa

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CXCL16 antibody [EPR29714-545] (AB324591)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CXCL16 with ab324591 at 1/50 (10.1 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on mouse cerebrum (PMID : 11017100).
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 Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-CXCL16 antibody [EPR29714-545] (AB324591)

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

CXCL16 undergoes cleavage and glycosylation modifications. Consequently, different band sizes corresponding to its distinct forms can be detected : the 28-37 kDa precursor form, the 50 kDa mature form, and the 38 kDa soluble form. Please refer to publications such as PMID : 15004171, PMID : 39436058, PMID : 33800554, PMID : 17467666.

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

All lanes:

Western blot - Anti-CXCL16 antibody [EPR29714-545] (ab324591) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a Myc-his tag, whole cell lysate at 10 µg

Lane 2:

293T cells transfected with a mouse CXCL16 expression vector containing a Myc-his tag, whole cell lysate at 10 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 1/100000 dilution

Observed band size: 50 kDa,38 kDa,28-37 kDa,20-50 kDa,36 kDa

false

Exposure time: 1s