WB
Western blot analysis of ALIX on different lysates with Rabbit anti-ALIX antibody at 1/2,000 dilution. Lane 1: HEK-293 cell lysate, Lane 2: K-562 cell lysate, Lane 3: Jurkat cell lysate, Lane 4: PC-3M cell lysate, Lane 5: MCF7 cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: 5 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.WB
Western blot analysis of ALIX on different lysates with Rabbit anti-ALIX antibody at 1/1,000 dilution. Lane 1: NIH/3T3 cell lysate (20 µg/Lane), Lane 2: PC-12 cell lysate (20 µg/Lane), Lane 3: HEK-293 cell lysate (20 µg/Lane), Lane 4: K-562 cell lysate (20 µg/Lane), Lane 5: Mouse liver tissue lysate (30 µg/Lane), Lane 6: Rat liver tissue lysate (30 µg/Lane), Exposure time: 6 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-ALIX antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of MCF7 cells labeling ALIX with Rabbit anti-ALIX antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ALIX antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of NIH/3T3 cells labeling ALIX. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | ALIX Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | ALG-2-interacting protein (Alix), also designated programmed cell death 6-interacting protein (PDCD6-interacting protein), is a cytoplasmic protein. Alix interacts with apoptosis-associated proteins (ALG-2 and PDCD6) and with the endocytosis-regulator CIN85. Additionally, Alix interacts with the endosomal sorting complexes required for transport (ESCRT) proteins (Tsg101 and CHMP4) and can associate with HIV-1. The endophilins (SH3P4, SH3P8 and SH3P13), enzymes that change curvature of the membrane that are required for early and late steps of coated vesicle formation, also bind to Alix. Alix is involved in the concentration and sorting of cargo proteins of the multivesicular body for incorporation into vesicles. |
| Immunogen | Recombinant protein within Human ALIX aa 1-160 / 868. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:1000-1:2000 IHC:1:50-1:200 ICC/IF:1:100 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | AIP1 antibody ALG 2 interacting protein 1 antibody ALG-2-interacting protein 1 antibody ALG2 interacting protein X antibody Alix antibody Apoptosis linked gene 2 interacting protein X antibody Dopamine receptor interacting protein 4 antibody DRIP4 antibody Hp95 antibody KIAA1375 antibody MGC17003 antibody PDC6I_HUMAN antibody PDCD6 interacting protein antibody PDCD6-interacting protein antibody PDCD6IP antibody Programmed cell death 6 interacting protein antibody Programmed cell death 6-interacting protein antibody |
|---|---|
| Molecular Weight(MW) | 96 kDa(Observed band size: 80-100 kDa) |
| Cellular Localization | Cell junction, Cytoplasm, Cytoskeleton, Secreted, Tight junction. |
WB
Western blot analysis of ALIX on different lysates with Rabbit anti-ALIX antibody at 1/2,000 dilution. Lane 1: HEK-293 cell lysate, Lane 2: K-562 cell lysate, Lane 3: Jurkat cell lysate, Lane 4: PC-3M cell lysate, Lane 5: MCF7 cell lysate, Lysates/proteins at 15 µg/Lane. Exposure time: 5 minutes; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.
WB
Western blot analysis of ALIX on different lysates with Rabbit anti-ALIX antibody at 1/1,000 dilution. Lane 1: NIH/3T3 cell lysate (20 µg/Lane), Lane 2: PC-12 cell lysate (20 µg/Lane), Lane 3: HEK-293 cell lysate (20 µg/Lane), Lane 4: K-562 cell lysate (20 µg/Lane), Lane 5: Mouse liver tissue lysate (30 µg/Lane), Lane 6: Rat liver tissue lysate (30 µg/Lane), Exposure time: 6 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-ALIX antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of MCF7 cells labeling ALIX with Rabbit anti-ALIX antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ALIX antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of NIH/3T3 cells labeling ALIX. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:1000-1:2000 IHC:1:50-1:200 ICC/IF:1:100 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
