Anti-Cystatin F antibody [EPR28408-30]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin F antibody [EPR28408-30] (AB309544)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Cystatin F with ab309544 at 1/100 (5.14 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : no staining on human cerebrum (PMID : 28178353). The section was incubated with ab309544 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin F antibody [EPR28408-30] (AB309544)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human tissue labeling Cystatin F with ab309544 at 1/100 (5.14 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on HEK-293T transfected with a His-tagged CST7 construct (image A). No staining on HEK-293T transfected with empty plasmid (image B). The section was incubated with ab309544 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

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cystatin F antibody [EPR28408-30] (AB309544)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Cystatin F with ab309544 at 1/100 (5.14 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human spleen. The section was incubated with ab309544 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Cystatin F antibody [EPR28408-30] (AB309544)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast, Left) / HL-60 (human acute promyelocytic leukemia promyeloblast, Right) cells labelling Cystatin F with ab309544 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Negative control : K-562.

• IP

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Immunoprecipitation - Anti-Cystatin F antibody [EPR28408-30] (AB309544)

Cystatin F was immunoprecipitated from 0.35 mg HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate with ab309544 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab309544 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate Lane 2 : ab309544 IP in HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab309544 in HL-60 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 3 seconds

All lanes:

Immunoprecipitation - Anti-Cystatin F antibody [EPR28408-30] (ab309544) at 1/30 dilution

All lanes:

HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell 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

false

Exposure time: 3s