Anti-MCP1 antibody

• WB

Supplier Data

Western blot - Anti-MCP1 antibody (AB25124)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 28955950).

Panel A is applied with Anti-MCP1 antibody [RM1100] (ab315478) at 1/1000 dilution and panel B is applied with Anti-MCP1 antibody (ab25124) at 1/1000 dilution.

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

Exposure time : 26 seconds (panel A);180 seconds (panel B).

All lanes:

Western blot - Anti-MCP1 antibody [RM1100] (<a href='/en-us/products/primary-antibodies/mcp1-antibody-rm1100-ab315478'>ab315478</a>) at 1/1000 dilution

Lane 1:

Untreated RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 treated with 100ng/ml Lipopolysaccharides (LPS) for 7 hours, then 1µg/ml Brefeldin A was added for additional 3 hours 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 1/100000 dilution

Observed band size: 20 kDa,36 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCP1 antibody (AB25124)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling MCP1 with ab315478 at 1/3000 (0.172 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : ab315478 showed no staining on mouse skeletal muscle (ab), while ab25124 showed non-specific staining on the mouse skeletal muscle.
The image on the left is applied with Anti-MCP1 antibody [RM1100] (ab315478) at 1/3000 dilution and the image on the right is applied with Anti-MCP1 antibody (ab25124) at 1/1500 dilution.
The section was incubated with ab315478 or ab25124 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