PA5-71659

antibody from Invitrogen Antibodies

Targeting: SMPD1 ASM

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-71659

Provider

Invitrogen Antibodies

Product name

ASM Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

400 µL

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of ASM in various lysates. Samples were incubated with ASM polyclonal antibody (Product # PA5-71659) using a dilution of 1:2,000 followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at a dilution of 1:10,000. Lysates/proteins: 20 µg per lane. Lane 1: human cerebellum lysate; Lane 2: HepG2 whole cell lysate; Lane 3: K562 whole cell lysate; Lane 4: MCF-7 whole cell lysate. Predicted band size: 70 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of ASM in various lysates. Samples were incubated with ASM polyclonal antibody (Product # PA5-71659) using a dilution of 1:4,000 followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at a dilution of 1:10,000. Lysates/proteins: 20 µg per lane. Lane 1: K562 whole cell lysate; Lane 2: MCF-7 whole cell lysate; Lane 3: HepG2 whole cell lysate; Lane 4: A431 whole cell lysate. Predicted band size: 70 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of ASM in paraffin-embedded Human testis carcinoma tissue. Samples were incubated with ASM polyclonal antibody (Product # PA5-71659) using a dilution of 1:500 for 1 hour at room temperature followed by an undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody. Tissue was fixed with formaldehyde at room temperature, antigen retrieval was by heat mediation with a EDTA buffer (pH 9.0).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry analysis of ASM in paraffin-embedded Human breast tissue. Samples were incubated with ASM polyclonal antibody (Product # PA5-71659) using a dilution of 1:500 for 1 hour at room temperature followed by an undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody. Tissue was fixed with formaldehyde at room temperature, antigen retrieval was by heat mediation with a EDTA buffer (pH 9.0).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of SMPD1 in K562 cells (green line). The cells were fixed with 2% paraformaldhyde (10 min) and permeabilized with 90% methanol (10 min). The sample was then incubated with 2% BSA followed by a SMPD1 polyclonal antibody (Product # PA5-71659) using a dilution of 1:25 for 60 min at 37°C. The secondary antibody used was goat-anti-rabbit IgG, DyLight 488 with a dilution of 1:200 for 40 min at 37°C. Isotype control antibody (blue line) was rabbit IgG (1 µg/1x10^6 cells) used under the same conditions with an acquisition of >10, 000 events.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of (overlay histogram) of ASM in K562 cells (green line). Samples were incubated with ASM polyclonal antibody (Product # PA5-71659) using a dilution of 1:25 dilution for 60 min at 37°C followed by Goat Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed at 1:200 dilution for 40 min at 37°C. The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then incubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the primary antibody. Isotype control antibody (blue line) was rabbit IgG (1 μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.