Anti-SDF1 antibody [EPR1216] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SDF1 antibody [EPR1216] - BSA and Azide free (AB271917)

Immunofluorescent analysis of Jurkat cells, labeling SDF1 with ab155090 at 1/100 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab155090).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SDF1 antibody [EPR1216] - BSA and Azide free (AB271917)

Immunocytochemistry/Immunofluorescence analysis of THP-1 (Human monocytic leukemia cell line) labeling SDF1 with Purified ab155090 at 1/500 dilution (5 μg/ml). Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab155090).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SDF1 antibody [EPR1216] - BSA and Azide free (AB271917)

Intracellular Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling SDF1 with purified ab155090 at 1/20 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab155090).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SDF1 antibody [EPR1216] - BSA and Azide free (AB271917)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Human PBMC (Human primary peripheral blood mononuclear cell) cells labelling SDF1 with primary antibody anti-SDF1 (ab155090) at 1/100 dilution, followed by Alexa Fluor^® 488 Goat anti-Rabbit secondary (ab150077) secondary antibody at 1/1000 dilution. Anti-alpha Tubulin antibody (DM1A) - Microtubule Marker (Alexa Fluor^® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution. The nuclear counter stain is DAPI (blue). Confocal image showing cytoplasmic staining in Human PBMC. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Low expression control : K562 (PMID : 23473997)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab155090).

• WB

Lab

Western blot - Anti-SDF1 antibody [EPR1216] - BSA and Azide free (AB271917)

This data was developed using ab155090, the same antibody clone in a different buffer formulation.

Western blot : Rabbit monoclonal [EPR1216] to SDF1 ab155090 staining at 1/500 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 25 kDa in Wild-type MCF7 serum starved (72 h), vehicle control EtOH (0.1%, last 48 h) cell lysates with no signal observed at this size in CXCL12 knockout MCF7 serum starved (72 h), treated E2 (10 nM, last 48 h) cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-SDF1 antibody [EPR1216] (<a href='/en-us/products/primary-antibodies/sdf1-antibody-epr1216-ab155090'>ab155090</a>) at 1/500 dilution

Lane 1:

Wild-type MCF7 serum starved (72 h), vehicle control EtOH (0.1%, last 48 h) at 20 µg

Lane 2:

Wild-type MCF7 serum starved (72 h), treated E2 (10 nM, last 48 h) at 20 µg

Lane 3:

CXCL12 knockout MCF7 serum starved (72 h), vehicle control EtOH (0.1%, last 48 h) at 20 µg

Lane 4:

CXCL12 knockout MCF7 serum starved (72 h), treated E2 (10 nM, last 48 h) at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 25 kDa

Observed band size: 25 kDa

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