Anti-SPARCL1 antibody [EPR28135-54] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPARCL1 antibody [EPR28135-54] - BSA and Azide free (AB313639)

This data was developed using ab313638, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling SPARCL1 with ab313638 at 1/2000 (0.248 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse cerebrum (PMID : 18381651).The section was incubated with ab313638 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background 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-SPARCL1 antibody [EPR28135-54] - BSA and Azide free (AB313639)

This data was developed using ab313638, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling SPARCL1 with ab313638 at 1/2000 (0.248 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on endothelium of mouse spleen (PMID : 9199668).The section was incubated with ab313638 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background 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

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-SPARCL1 antibody [EPR28135-54] - BSA and Azide free (AB313639)

This data was developed using ab313638, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling SPARCL1 with ab313638 at 1/50 (9.9 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing cytoplasmic staining in mouse astrocyte cells.Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. Anti-Glial Fibrillary Acidic Protein (GFAP) mouse monoclonal antibody was used to counterstain tubulin at 1/50 10ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SPARCL1 antibody [EPR28135-54] - BSA and Azide free (AB313639)

This data was developed using ab313638, the same antibody clone in a different buffer formulation. Flow cytometric analysis of Mouse primary neural/glia cell cells labelling SPARCL1 with ab313638 at 1/50 dilution (1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-SPARCL1 antibody [EPR28135-54] - BSA and Azide free (AB313639)

This data was developed using ab313638, the same antibody clone in a different buffer formulation. SPARCL1 was immunoprecipitated from 0.35 mg Mouse cerebellum tissue lysate with ab313638 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313638 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse cerebellum tissue lysate Lane 2 : ab313638 IP in Mouse cerebellum tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313638 in mouse cerebellum tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 10 seconds

All lanes:

Immunoprecipitation - Anti-SPARCL1 antibody [EPR28135-54] (<a href='/en-us/products/primary-antibodies/sparcl1-antibody-epr28135-54-ab313638'>ab313638</a>) at 1/30 dilution

All lanes:

Mouse cerebellum tissue 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 1/1000 dilution

false

Exposure time: 10s

• WB

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Western blot - Anti-SPARCL1 antibody [EPR28135-54] - BSA and Azide free (AB313639)

This data was developed using ab313638, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The molecular weight observed is consistent with what has been described in the literature (PMID : 22173850). Low expression : kidney (PMID : 9199668; PMID : 32437418). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 6 seconds

All lanes:

Western blot - Anti-SPARCL1 antibody [EPR28135-54] (<a href='/en-us/products/primary-antibodies/sparcl1-antibody-epr28135-54-ab313638'>ab313638</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse cerebellum tissue lysate at 20 µg

Lane 3:

Mouse hippocampus tissue lysate at 20 µg

Lane 4:

Mouse lung tissue lysate at 20 µg

Lane 5:

Mouse kidney tissue 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 1/20000 dilution

Observed band size: 130 kDa

false

Exposure time: 6s