HA723797

Perforin-1 Recombinant Rabbit Monoclonal Antibody [PSH15-92]

cat.: HA723797

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P
Clonality:	Monoclonal
Clone number:	PSH15-92

Form:	Liquid
Storage condition:	Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 61 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within mouse Perforin-1 aa 351-555.
Positive control:	CTLL-2 cell lysate, NK-92 cell lysate, rat spleen lysates, rat spleen tissue, rat lung tissue, CTLL-2.
Subcellular location:	Cytoplasm,Cell membrane,extracellular region.
Recommended Dilutions: WB IF-Cell IHC-P	1:5,000-1:20,000 1:500 1:200-1:1,000
Uniprot #:	SwissProt: P14222 Human \| P10820 Mouse \| P35763 Rat
Alternative names:	Cytolysin FLH2 HPLH2 Lymphocyte pore-forming protein P1 PERF_HUMAN perforin 1 (pore forming protein) Perforin 1 Perforin-1 PFP PGFL PIGF PIGF-2 PLGF Pore forming protein prf1 SHGC-10760

Images

Fig1: Western blot analysis of Perforin-1 on different lysates with Rabbit anti-Perforin-1 antibody (HA723797) at 1/20,000 dilution.

Lane 1: NIH/3T3 cell lysate (negative)
Lane 2: CTLL-2 cell lysate
Lane 3: C2C12 cell lysate (negative)

Lysates/proteins at 10 µg/Lane.

Predicted band size: 61 kDa
Observed band size: 75 kDa

Exposure time: 4 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 (HA723797) at 1/20,000 dilution was used in primary antibody dilution (K1803) 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 Perforin-1 on different lysates with Rabbit anti-Perforin-1 antibody (HA723797) at 1/10,000 dilution.

Lane 1: Ramos cell lysate (negative)
Lane 2: NK-92 cell lysate
Lane 3: 293T cell lysate (negative)

Lysates/proteins at 20 µg/Lane.

Predicted band size: 61 kDa
Observed band size: 61 kDa

Exposure time: 3 minutes; 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 (HA723797) at 1/10,000 dilution was used in primary antibody dilution (K1803) 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: Western blot analysis of Perforin-1 on rat spleen tissue lysates with Rabbit anti-Perforin-1 antibody (HA723797) at 1/5,000 dilution.

Lysates/proteins at 40 µg/Lane.

Predicted band size: 61 kDa
Observed band size: 61 kDa

Exposure time: 25 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 (HA723797) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig4: Immunocytochemistry analysis of CTLL-2 (positive) and NIH/3T3 (negative) labeling Perforin-1 with Rabbit anti-Perforin-1 antibody (HA723797) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-Perforin-1 antibody (HA723797) at 1/500 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 (HA601187, 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.

Fig5: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Prf1 antibody (HA723797) 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 (HA723797) 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: Immunohistochemical analysis of paraffin-embedded rat lung tissue with Rabbit anti-Prf1 antibody (HA723797) 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 (HA723797) 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.

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