ET1704-90

Calpain 2 Recombinant Rabbit Monoclonal Antibody [JA43-41]

cat.: ET1704-90

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JA43-41

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: 80 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human Calpain 2 aa 651-700 / 700.
Positive control:	HCT 116 cell lysate, MDA-MB-231 cell lysate, 293T cell lysate, HepG2, human lung tissue, human kidney tissue, Hela.
Subcellular location:	Cytoplasm. Cell membrane.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:500-1:2,000 1:50 1:50-1:200 1:50
Uniprot #:	SwissProt: P17655 Human \| O08529 Mouse \| Q07009 Rat
Alternative names:	Calcium activated neutral proteinase 2 Calcium activated neutral proteinase Calcium-activated neutral protease 2, catalytic subunit Calcium-activated neutral proteinase 2 CALP80 Calpain 2 (m/II) large subunit Calpain 2 catalytic subunit Calpain 2 large catalytic subunit Calpain 2 large subunit Calpain 2, large [catalytic] subunit Calpain large polypeptide L2 Calpain M type Calpain M-type Calpain-2 catalytic subunit Calpain-2 large subunit Calpain2 CAN2_HUMAN CANP 2 CANP L2 CANP2 CANPL 2 CANPL2 CANPml Capa2 CAPN 2 CAPN2 FLJ39928 M calpain M calpin M type M-calpain mCANP Millimolar calpain Millimolar-calpain

Images

Fig1: Western blot analysis of Calpain 2 on different lysates with Rabbit anti-Calpain 2 antibody (ET1704-90) at 1/1,000 dilution.

Lane 1: HCT 116-si NT cell lysate
Lane 2: HCT 116-si Calpain 2 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 80 kDa
Observed band size: 75 kDa

Exposure time: 40 seconds; ECL: K1801;

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 (ET1704-90) at 1/1,000 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.

Fig2: Western blot analysis of Calpain 2 on different lysates with Rabbit anti-Calpain 2 antibody (ET1704-90) at 1/1,000 dilution.

Lane 1: MDA-MB-231 cell lysate
Lane 2: 293T cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 80 kDa
Observed band size: 70 kDa

Exposure time: 1 minutes; ECL: K1802;

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 (ET1704-90) at 1/1,000 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: ICC staining of Calpain 2 in HepG2 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 (ET1704-90, 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).
	Fig4: Immunohistochemical analysis of paraffin-embedded human lung tissue using anti-Calpain 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 (ET1704-90, 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 kidney tissue with Rabbit anti-Calpain 2 antibody (ET1704-90) 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 (ET1704-90) 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.
	Fig6: Flow cytometric analysis of Calpain 2 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1704-90, 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 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.