Integrin alpha V Recombinant Rabbit Monoclonal Antibody [SC56-07]
cat.: ET1610-15
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC, IHC-P
Clonality: Monoclonal
Clone number: SC56-07
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 116 kDa
Isotype: IgG
Immunogen: Synthetic peptide within of Human Integrin alpha V aa 1000-1048 / 1048.
Positive control: A549 cell lysate, HeLa cell lysate, NIH/3T3 cell lysate, C6 cell lysate, rat heart tissue lysate, A549, SiHa, MCF-7, human placenta tissue.
Subcellular location: Membrane.
Recommended Dilutions:
  WB
  IF-Cell
  FC
  IHC-P
1:500-1:5,000
1:50-1:100
1:50-1:100
1:1,000
Uniprot #: SwissProt: P06756 Human | P43406 Mouse
Entrez Gene: 296456 Rat
Alternative names: CD51 Integrin alpha-V light chain integrin, alpha V (vitronectin receptor, alpha polypeptide, antigen CD51) Integrin, alpha V Integrin, beta 6 ITAV_HUMAN ITGAV ITGB6 Vitronectin receptor subunit alpha
Images
ET1610-15_1.jpg Fig1: Western blot analysis of Integrin alpha V on different lysates with Rabbit anti-Integrin alpha V antibody (ET1610-15) at 1/2,000 dilution.

Lane 1: A549 cell lysate (10 µg/Lane)
Lane 2: HeLa cell lysate (10 µg/Lane)
Lane 3: NIH/3T3 cell lysate (10 µg/Lane)
Lane 4: C6 cell lysate (10 µg/Lane)
Lane 5: Rat heart tissue lysate (20 µg/Lane)

Predicted band size: 116 kDa
Observed band size: 125/135 kDa

Exposure time: 25 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 (ET1610-15) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1610-15_2.jpg Fig2: All lanes: Western blot analysis of Integrin alpha V with anti-Integrin alpha V antibody (ET1610-15) at 1/500 dilution.

Lane 1: Wild-type A549 whole cell lysate.
Lane 2/3: Integrin alpha V knockdown A549 whole cell lysate.

ET1610-15 was shown to specifically react with Integrin alpha V in Wild-type A549 cells. Weakened bands were observed when Integrin alpha V knockdown samples were tested. Wild-type and Integrin alpha V knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-Integrin alpha V antibody (ET1610-15, 1/500) and Anti-HSP90 antibody (ET1605-56, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
ET1610-15_3.jpg Fig3: Western blot analysis of Integrin alpha V on different lysates with Rabbit anti-Integrin alpha V antibody (ET1610-15) at 1/5,000 dilution.

Lane 1: A549 cell lysate
Lane 2: HeLa cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 116 kDa
Observed band size: 130 kDa

Exposure time: 4 minutes;

6% 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 (ET1610-15) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
ET1610-15_4.jpg Fig4: ICC staining of Integrin alpha V in A549 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-15, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1610-15_5.jpg Fig5: ICC staining of Integrin alpha V in SiHa cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-15, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1610-15_6.jpg Fig6: Flow cytometric analysis of Integrin alpha V was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1610-15, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1610-15_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-Integrin alpha V antibody (ET1610-15) at 1/1,000 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 (ET1610-15) at 1/1,000 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.