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

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Specification

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Catalog# HA722222

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

Application

  • WB

  • IF-Cell

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

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

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Human Caspase-1 aa 255-404 / 404.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P

Target Molecular Weight

Predicted band size: 45 kDa

Positive Control

Raji cell lysate, THP-1 cell lysate, U-937 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.

Conjugation

unconjugated

Clone Number

JE56-35

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000-1:5,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:500-1:2,000

Target

Function

Caspase-1/Interleukin-1 converting enzyme (ICE) is an evolutionarily conserved enzyme that proteolytically cleaves other proteins, such as the precursors of the inflammatory cytokines interleukin 1β and interleukin 18 as well as the pyroptosis inducer Gasdermin D, into active mature peptides. It plays a central role in cell immunity as an inflammatory response initiator. Once activated through formation of an inflammasome complex, it initiates a proinflammatory response through the cleavage and thus activation of the two inflammatory cytokines, interleukin 1β (IL-1β) and interleukin 18 (IL-18) as well as pyroptosis, a programmed lytic cell death pathway, through cleavage of Gasdermin D. The two inflammatory cytokines activated by Caspase-1 are excreted from the cell to further induce the inflammatory response in neighboring cells.

Background References

1. Wang F et al. Silica nanoparticles induce pyroptosis and cardiac hypertrophy via ROS/NLRP3/Caspase-1 pathway. Free Radic Biol Med. 2022 Mar

2. Kang L et al. Blocking Caspase-1/Gsdmd and Caspase-3/-8/Gsdme pyroptotic pathways rescues silicosis in mice. PLoS Genet. 2022 Dec

Subcellular Location

Cytoplasm, Cell membrane, nucleus.

UNIPROT

SwissProt: P29466 Human

SwissProt: P29452 Mouse

SwissProt: P43527 Rat

Synonyms

CASP1 antibody

Caspase 1 apoptosis related cysteine peptidase antibody

ICE antibody

IL 1 beta converting enzyme antibody

IL 1BC antibody

IL1B convertase antibody

Interleukin 1 beta convertase antibody

Interleukin 1 beta converting enzyme antibody

p45 antibody

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of Caspase-1 + p10 + p12 on different lysates with Rabbit anti-Caspase-1 + p10 + p12 antibody (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) at 1/5,000 dilution.<br /><br />Lane 1: Raji cell lysate<br />Lane 2: THP-1 cell lysate<br />Lane 3: U-937 cell lysate<br />Lane 4: A549 cell lysate (low expression)<br />Lane 5: RAW264.7 cell lysate<br />Lane 6: Mouse spleen tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 45 kDa<br />Observed band size: 45/42/35/12/10 kDa<br /><br />Exposure time: 10 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of Caspase-1 + p10 + p12 on different lysates with Rabbit anti-Caspase-1 + p10 + p12 antibody (HA722222) at 1/5,000 dilution.

    Lane 1: Raji cell lysate
    Lane 2: THP-1 cell lysate
    Lane 3: U-937 cell lysate
    Lane 4: A549 cell lysate (low expression)
    Lane 5: RAW264.7 cell lysate
    Lane 6: Mouse spleen tissue lysate

    Lysates/proteins at 20 µg/Lane.

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

    Exposure time: 10 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 1/5,000 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.

  • Western blot analysis of Caspase-1 + p10 + p12 on rat spleen tissue lysates with Rabbit anti-Caspase-1 + p10 + p12 antibody (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) at 1/5,000 dilution.<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 45 kDa<br />Observed band size: 45/12/10 kDa<br /><br />Exposure time: 3 minute 30 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) at 1/5,000 dilution was used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Caspase-1 + p10 + p12 on rat spleen tissue lysates with Rabbit anti-Caspase-1 + p10 + p12 antibody (HA722222) at 1/5,000 dilution.

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 3 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 1/5,000 dilution was used in 5% BSA at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of THP-1 cells labeling Caspase-1 + p10 + p12 with Rabbit anti-Caspase-1 + p10 + p12 antibody (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    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.

  • Immunocytochemistry analysis of RAW264.7 cells labeling Caspase-1 + p10 + p12 with Rabbit anti-Caspase-1 + p10 + p12 antibody (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    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.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-Caspase-1 + p10 + p12 antibody (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Caspase-1 + p10 + p12 antibody (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Caspase-1 + p10 + p12 antibody (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) at 1/2,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722222" style="font-weight: bold;text-decoration: underline;">HA722222</a>) 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.

    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.

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Citation

Products with the same target and pathway

metafield image

Caspase-1 + p10 + p12 Recombinant Rabbit Monoclonal Antibody [JE56-35] - BSA and Azide free

Application: WB,IF-Cell,IHC-P

Reactivity: Human,Mouse,Rat

Conjugate: unconjugated