CCL3 / MIP-1 alpha Recombinant Rabbit Monoclonal Antibody [PSH11-28]

☑ Cell treatment (CT)

Western blot analysis of CCL3 / MIP-1 alpha on different lysates with Rabbit anti-CCL3 / MIP-1 alpha antibody (HA723310) at 1/2,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: THP-1 treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours then add 1μg/mL BFA for 3 hours cell lysate
Lane 3: RAW264.7 cell lysate
Lane 4: RAW264.7 treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours then add 1μg/mL BFA for 3 hours cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 10 kDa
Observed band size: 10 kDa

Exposure time: 25 seconds; ECL: K1802;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723310) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Relative expression (RE)

Western blot analysis of CCL3 / MIP-1 alpha on different lysates with Rabbit anti-CCL3 / MIP-1 alpha antibody (HA723310) at 1/2,000 dilution.

Lane 1: NK-92 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (negative) (20 µg/Lane)

Predicted band size: 10 kDa
Observed band size: 10 kDa

Exposure time: 25 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723310) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Cell treatment (CT)

Immunocytochemistry analysis of RAW264.7 cells untreated / treated with 100nM TPA overnight then add 100ng/mL LPS for 7 hours labeling CCL3 / MIP-1 alpha with Rabbit anti-CCL3 / MIP-1 alpha antibody (HA723310) at 1/2,000 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CCL3 / MIP-1 alpha antibody (HA723310) at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Immunohistochemical analysis of paraffin-embedded human hodgkin lymphoma tissue with Rabbit anti-CCL3 / MIP-1 alpha antibody (HA723310) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA723310) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CCL3 / MIP-1 alpha antibody (HA723310) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA723310) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.