HA722222

Caspase-1 + p10 + p12 Recombinant Rabbit Monoclonal Antibody [JE56-35]

cat.: HA722222

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P
Clonality:	Monoclonal
Clone number:	JE56-35

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. 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: 45/42/35/12/10 kDa
Isotype:	IgG
Immunogen:	Recombinant protein.
Positive control:	Raji cell lysate, THP-1 cell lysate, U-937 cell lysate, HepG2 cell lysate, RAW264.7 cell lysate, mouse spleen tissue lysate, rat spleen tissue lysate, THP-1, RAW264.7, human spleen tissue, mouse spleen tissue, rat spleen tissue.
Subcellular location:	Cytoplasm, Cell membrane.
Recommended Dilutions: WB IF-Cell IHC-P	1:2,000 1:100 1:500-1:2,000
Uniprot #:	SwissProt: P29466 Human \| P29452 Mouse \| P43527 Rat
Alternative names:	CASP1 Caspase 1 apoptosis related cysteine peptidase ICE IL 1 beta converting enzyme IL 1BC IL1B convertase Interleukin 1 beta convertase Interleukin 1 beta converting enzyme p45

Images

Fig1: Western blot analysis of Caspase-1 + p10 + p12 on different lysates with Rabbit anti-Caspase-1 + p10 + p12 antibody (HA722222) at 1/2,000 dilution.

Lane 1: Raji cell lysate
Lane 2: THP-1 cell lysate
Lane 3: U-937 cell lysate
Lane 4: HepG2 cell lysate
Lane 5: RAW264.7 cell lysate
Lane 6: Mouse spleen tissue lysate
Lane 7: Rat spleen tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 45/42/35/12/10 kDa
Observed band size: 45/42/35/12/10 kDa

Exposure time: 1 minute 30 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 (HA722222) 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.

Fig2: Immunocytochemistry analysis of THP-1 cells labeling Caspase-1 + p10 + p12 with Rabbit anti-Caspase-1 + p10 + p12 antibody (HA722222) 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-Caspase-1 + p10 + p12 antibody (HA722222) 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.

	Fig3: Immunocytochemistry analysis of RAW264.7 cells labeling Caspase-1 + p10 + p12 with Rabbit anti-Caspase-1 + p10 + p12 antibody (HA722222) 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-Caspase-1 + p10 + p12 antibody (HA722222) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Caspase-1 + p10 + p12 antibody (HA722222) at 1/2,000 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 (HA722222) at 1/2,000 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Caspase-1 + p10 + p12 antibody (HA722222) at 1/2,000 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 (HA722222) at 1/2,000 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.

Fig6: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Caspase-1 + p10 + p12 antibody (HA722222) at 1/2,000 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 (HA722222) at 1/2,000 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”.