Anti-Caspase-3 antibody [RM1299] (ab323369)

Rabbit Recombinant Multiclonal Caspase-3 antibody. Suitable for WB, IHC-P, IP and reacts with Human, Mouse samples.

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Images

Immunoprecipitation - Anti-Caspase-3 antibody [RM1299] (AB323369), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Multiclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-P	IP	Flow Cyt (Intra)	ICC/IF
Human	Tested	Tested	Tested	Not recommended	Not recommended
Mouse	Tested	Tested	Tested	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -
Species Mouse	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/5000	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/5000	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes -
Species Mouse	Dilution info 1/30	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Mouse, Rat	Dilution info -	Notes -

Target data

See full target information Casp3

Function

Thiol protease that acts as a major effector caspase involved in the execution phase of apoptosis (PubMed:16469926, PubMed:8934524). Following cleavage and activation by initiator caspases (CASP8, CASP9 and/or CASP10), mediates execution of apoptosis by catalyzing cleavage of many proteins (PubMed:16469926, PubMed:8934524). At the onset of apoptosis, it proteolytically cleaves poly(ADP-ribose) polymerase PARP1 at a '216-Asp-|-Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9 (CASP6, CASP7 and CASP9, respectively). Cleaves and inactivates interleukin-18 (IL18) (By similarity). Triggers cell adhesion in sympathetic neurons through RET cleavage (By similarity). Cleaves IL-1 beta between an Asp and an Ala, releasing the mature cytokine which is involved in a variety of inflammatory processes (By similarity). Cleaves and inhibits serine/threonine-protein kinase AKT1 in response to oxidative stress (PubMed:12124386). Acts as an inhibitor of type I interferon production during virus-induced apoptosis by mediating cleavage of antiviral proteins CGAS, IRF3 and MAVS, thereby preventing cytokine overproduction (PubMed:30878284). Also involved in pyroptosis by mediating cleavage and activation of gasdermin-E (GSDME) (By similarity). Cleaves XRCC4 and phospholipid scramblase proteins XKR4, XKR8 and XKR9, leading to promote phosphatidylserine exposure on apoptotic cell surface (PubMed:25231987, PubMed:33725486). Cleaves BIRC6 following inhibition of BIRC6-caspase binding by DIABLO/SMAC (By similarity).

Targets

CASP3

Alternative names

Cpp32, Casp3, Caspase-3, CASP-3, Apopain, Cysteine protease CPP32, LICE, Protein Yama, SREBP cleavage activity 1, CPP-32, SCA-1

Rabbit Recombinant Multiclonal Caspase-3 antibody. Suitable for WB, IHC-P, IP and reacts with Human, Mouse samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Multiclonal
Immunogens: The exact immunogen used to generate this antibody is proprietary information.
Clone number: RM1299
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant multiclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

Immunoprecipitation - Anti-Caspase-3 antibody [RM1299] (ab323369)
Caspase-3 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab323369 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323369 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Blocking and dilution buffer and concentration: 5% NFDM/TBST
All lanes: Immunoprecipitation - Anti-Caspase-3 antibody [RM1299] (ab323369) at 1/1000 dilution
Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 10 µg
Lane 2: ab323369 at 1/30 IP in NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab323369 in NIH/3T3 whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 32s
Immunoprecipitation - Anti-Caspase-3 antibody [RM1299] (ab323369)
Caspase-3 was immunoprecipitated from 0.35 mg parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate with ab323369 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323369 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Blocking and dilution buffer and concentration: 5% NFDM/TBST
All lanes: Immunoprecipitation - Anti-Caspase-3 antibody [RM1299] (ab323369) at 1/1000 dilution
Lane 1: parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate at 10 µg
Lane 2: ab323369 at 1/30 IP in parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell ) whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab323369 in parental HAP1(HOWT01) whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 6s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [RM1299] (ab323369)
Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling Caspase-3 with ab323369 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Low expression: nearly no staining on mouse cardiac muscle.

The section was incubated with ab323369 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [RM1299] (ab323369)
Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling Caspase-3 with ab323369 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Low expression: no staining on human skeletal muscle.

The section was incubated with ab323369 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [RM1299] (ab323369)
Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Caspase-3 with ab323369 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on mouse colon.

The section was incubated with ab323369 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [RM1299] (ab323369)
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Caspase-3 with ab323369 at 1/5000 (0.10 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human tonsil.

The section was incubated with ab323369 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins
Western blot - Anti-Caspase-3 antibody [RM1299] (ab323369)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 11423986).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-Caspase-3 antibody [RM1299] (ab323369) at 1/1000 dilution
Lane 1: Untreated NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 2: NIH/3T3 treated with 2uM staurosporine for 6 hours, whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 32 kDa, 17 kDa, 36 kDa
Exposure time: 6s
Western blot - Anti-Caspase-3 antibody [RM1299] (ab323369)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 11423986).
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
Exposure time: Lane 1-2: 70 seconds; Lane 3-4: 15 seconds
All lanes: Western blot - Anti-Caspase-3 antibody [RM1299] (ab323369) at 1/1000 dilution
Lane 1: Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: HeLa treated with 2uM staurosporine for 3 hours, whole cell lysate at 20 µg
Lane 3: Untreated Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg
Lane 4: Jurkat treated with 1uM staurosporine for 3 hours, whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 32 kDa, 17 kDa, 36 kDa
Western blot - Anti-Caspase-3 antibody [RM1299] (ab323369)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: skeletal muscle
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-Caspase-3 antibody [RM1299] (ab323369) at 1/1000 dilution
Lane 1: Human tonsil tissue lysate at 20 µg
Lane 2: Human skeletal muscle tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 32 kDa, 36 kDa
Exposure time: 15s
Western blot - Anti-Caspase-3 antibody [RM1299] (ab323369)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: heart
In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.
All lanes: Western blot - Anti-Caspase-3 antibody [RM1299] (ab323369) at 1/1000 dilution
Lane 1: Human stomach tissue lysate at 20 µg
Lane 2: Human spleen tissue lysate at 20 µg
Lane 3: Human testis tissue lysate at 20 µg
Lane 4: Mouse lung tissue at 20 µg
Lane 5: Mouse spleen tissue lysate at 20 µg
Lane 6: Mouse heart tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 32 kDa, 36 kDa
Exposure time: 37s
Western blot - Anti-Caspase-3 antibody [RM1299] (ab323369)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
In Western blot, ab323369 was shown to bind specifically to Caspase-3. Target of interest was observed at 32 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in Caspase-3 knockout cell line (lane 2).
In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842) staining at 1/100000 dilution.
All lanes: Western blot - Anti-Caspase-3 antibody [RM1299] (ab323369) at 1/1000 dilution
Lane 1: Parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg
Lane 2: Caspase-3 knockout HAP1 whole cell lysate at 20 µg
Lane 3: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4: Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg
Lane 5: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 6: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg
Lane 7: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 32 kDa, 15 kDa
Exposure time: 26s