ET1611-57

Integrin alpha 2 Recombinant Rabbit Monoclonal Antibody [SN0752]

cat.: ET1611-57

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	SN0752

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: 129 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human Integrin alpha 2 aa 1,132-1,181 / 1,181.
Positive control:	MCF7 cell lysate, mouse spleen tissue lysate, rat spleen tissue lysate, human colon carcinoma tissue, human breast carcinoma tissue, human kidney tissue, mouse colon tissue, mouse stomach tissue, A431.
Subcellular location:	Membrane.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:5,000 1:100-1:500 1:100-1:500 1:50-1:200 1:50-1:100
Uniprot #:	SwissProt: P17301 Human \| Q62469 Mouse Unigene: 83597 Rat
Alternative names:	BR CD 49b CD49 antigen like family member B CD49 antigen-like family member B CD49b CD49b antigen Collagen receptor DX5 Glycoprotein Ia deficiency included GP Ia GP Ia deficiency, included GPIa HPA 5 included HPA5 included Human platelet alloantigen system 5 Integrin alpha 2 Integrin alpha-2 Integrin, alpha 2 (CD49B alpha 2 subunit of VLA 2 receptor) ITA2_HUMAN ITGA2 Platelet alloantigen Br(a), included Platelet antigen Br Platelet glycoprotein GPIa Platelet glycoprotein Ia Platelet glycoprotein Ia/IIa Platelet membrane glycoprotein Ia Platelet receptor for collagen, deficiency of, included Very late activation protein 2 receptor alpha 2 subunit VLA 2 alpha chain VLA 2 VLA 2 subunit alpha VLA-2 subunit alpha VLA2 VLA2 receptor alpha 2 subunit VLAA2

Images

Fig1: Western blot analysis of Integrin alpha 2 with anti-Integrin alpha 2 antibody [SN0752] (ET1611-57) at 1/1,000 dilution.
Lane 1/2: Wild-type Hela whole cell lysate (20 µg).
Lane 3/4: Integrin alpha 2 fragment 1 knockdown Hela whole cell lysate (20 µg).
Lane 5/6: Integrin alpha 2 fragment 2 knockdown Hela whole cell lysate (20 µg).

ET1611-57 was shown to specifically react with Integrin alpha 2 in wild-type Hela cells. Weakened bands were observed when Integrin alpha 2 knockdown samples were tested. Wild-type and Integrin alpha 2 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 antibody (ET1611-57, 1/1,000) and Loading control antibody (Rabbit anti-HSP90, ET1605-56, 1/10,000) were used in 5% BSA 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.

Fig2: Western blot analysis of Integrin alpha 2 on different lysates with Rabbit anti-Integrin alpha 2 antibody (ET1611-57) at 1/5,000 dilution.

Lane 1: MCF7 cell lysate
Lane 2: Mouse spleen tissue lysate
Lane 3: Rat spleen tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 129 kDa
Observed band size: 150 kDa

Exposure time: 24 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 (ET1611-57) at 1/5,000 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.

	Fig3: Integrin alpha 2 was immunoprecipitated in 0.2mg MCF7 cell lysate with ET1611-57 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1611-57 at 1/2,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: MCF7 cell lysate (input) Lane 2: Rabbit IgG instead of ET1611-57 in MCF7 cell lysate Lane 3: ET1611-57 IP in MCF7 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 43 seconds
	Fig4: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Integrin alpha 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-57, 1/50) for 30 minutes 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Integrin alpha 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-57, 1/50) for 30 minutes 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Integrin alpha 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-57, 1/50) for 30 minutes 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.

	Fig7: Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-Integrin alpha 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-57, 1/50) for 30 minutes 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded mouse stomach tissue using anti-Integrin alpha 2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1611-57, 1/50) for 30 minutes 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.
	Fig9: Flow cytometric analysis of Integrin alpha 2 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1611-57, 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).
	Fig10: Immunocytochemistry analysis of A431 cells labeling Integrin alpha 2 with Rabbit anti-Integrin alpha 2 antibody (ET1611-57) 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-Integrin alpha 2 antibody (ET1611-57) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution."

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