PA5-19546

antibody from Invitrogen Antibodies

Targeting: KHDRBS1 FLJ34027, p62, Sam68

Immunocytochemistry

Immunohistochemistry

Antibody data

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Validation data

Product number: PA5-19546 - Provider product page
Provider: Invitrogen Antibodies
Product name: Anti-SAM68 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: Heat mediated antigen retrieval recommended prior to tissue staining. This antibody is predicted to react with chicken and dog based on sequence homology.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 1 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

KHDRBS1 protein structure - PA5-19546 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of SAM68 was performed by loading 10 µg of whole cell human BJ fibroblast protein lysate and run on a 4-12% BTE gel. Proteins were transferred to PVDF membrane. Membrane was blocked with Pierce™ Protein-Free T20 (PBS) Blocking Buffer (Product # 37573). SAM68 was detected at approximately 68 kDa using a SAM68 polyclonal antibody (Product # PA5-19546) at a dilution of 1:1000 followed by a 1:10,000 dilution of anti-rabbit HRP. Chemiluminescent detection was performed using SuperSignal West Pico PLUS substrate (Product # 34580). Data courtesy of Antibody Data Exchange Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HeLa Whole Cell Lysate using Product # PA5-19546, SAM68 primary antibody at a dilution of 1 µg/mL (lane 1). Staining of Jurkat Whole Cell Lysate at a dilution of 1 µg/mL (lane 2). Blot treated with a secondary HRP-conjugated Goat polyclonal anti-Rabbit antibody was used at a dilution of 1:3000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on modified whole cell extracts (1% SDS) (30 µg lysate) of MCF7 (Lane 1), MCF7 treated with Insulin (1nM for 24hr) (Lane 2), HeLa (Lane 3), Jurkat (Lane 4), A-431 (Lane 5) and U-87 MG (Lane 6). The blot was probed with Anti-SAM68 Polyclonal Antibody (Product # PA5-19546, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 57 kDa band corresponding to SAM68 was detected across the cell lines tested and the expression was also found to increase upon Insulin treatment in MCF7 cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of SAM68 in human fibroblasts. Cells were fixed with 2% PFA in PBS and permeabilized with 0.1% Triton X-100 in PBS. Cells were blocked with 3% Donkey serum in PBS followed by staining with a SAM68 polyclonal antibody (Product # PA5-19546) at a dilution of 1:200. Cells washed extensively followed by an incubation with anti-rabbit Alexa Fluor 594 (Product # A-21207) at a dilution of 1:500 (red). Data courtesy of Antibody Data Exchange Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent staining of HEK293 cells using Product # PA5-19546, anti-SAM68 antibody. The cells were fixed with methanol (100%) for 5 minutes, permabilised with TBS-T (20mins), BSA (1%), normal goat serum (10%) and glycine (0.3 M) in 0.1% PBS-Tween for 1 hour and exposed to the primary antibody at a concentration of 1 µg/mL for 1 hour at room temp. The secondary antibody was a 448 fluorescence conjugated Goat anti-rabbit IgG (green) at a dilution of 1:1000. A WGA- 594 fluorescent conjugated stain was used to label plasma membranes (red) and the nuclei stain was DAPI (blue).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of SAM68 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with SAM68 Polyclonal Antibody (Product # PA5-19546) at 1µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.