Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR3324] to Calpain small subunit 1
- Suitable for: WB, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Rat, Human
Product nameAnti-Calpain small subunit 1 antibody [EPR3324]
See all Calpain small subunit 1 primary antibodies
DescriptionRabbit monoclonal [EPR3324] to Calpain small subunit 1
Tested applicationsSuitable for: WB, IHC-P, Flow Cytmore details
Unsuitable for: IP
Species reactivityReacts with: Mouse, Rat, Human
Predicted to work with: Chinese hamster
Synthetic peptide within Human Calpain small subunit 1 aa 100-200. The exact sequence is proprietary.
- WB: fetal brain or fetal spleen tissue lysate; T47D or 293T cell lysate. IHC: Human kidney tissue.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
Concentration information loading...
PurityTissue culture supernatant
Our Abpromise guarantee covers the use of ab92333 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ICC: 1/100 - 1/250.
IHC-P: 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. The use of an HRP/AP polymerized antibody will give a stronger signal.
WB: 1/1000 - 1/5000. Predicted molecular weight: 28 kDa.
Is unsuitable for IP.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
FunctionRegulatory subunit of the calcium-regulated non-lysosomal thiol-protease which catalyzes limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction.
Sequence similaritiesContains 5 EF-hand domains.
DomainThe contact of the 5th EF-hand domain from each monomer allows the formation of the homodimer and also appears to mediate the contact between the large catalytic subunit and small regulatory subunit for the formation of the heterodimer.
EF-hand domains are paired. EF-hand 1 is paired with EF-hand 2 and EF-hand 3 is paired with EF-hand 4. The fifth EF-hand domain, left unpaired, does not bind the calcium but is responsible of the dimerization by EF-embrace. The first four EF-hand domains bind calcium, however it is not sure if the binding of EF-hand 4 to calcium is physiologically relevant.
Cellular localizationCytoplasm. Cell membrane. Translocates to the plasma membrane upon calcium binding.
- Information by UniProt
- 30K antibody
- Calcium activated neutral proteinase antibody
- Calcium activated neutral proteinase small subunit antibody
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Calpain small subunit 1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: A431 whole cell lysate (20 µg)
Lane 4: HEK293 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab92333 observed at 28 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab92333 was shown to recognize Calpain small subunit 1 when Calpain small subunit 1 knockout samples were used, along with additional cross-reactive bands. Wild-type and Calpain small subunit 1 knockout samples were subjected to SDS-PAGE. Ab92333 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes : Anti-Calpain small subunit 1 antibody [EPR3324] (ab92333) at 1/2000 dilution
Lane 1 : T47D
Lane 2 : fetal brain lysate
Lane 3 : fetal spleen lysate
Lane 4 : 293T
Lysates/proteins at 10 µg per lane.
All lanes : Goat anti-Rabbit HRP at 1/2000 dilution
Predicted band size: 28 kDa
Observed band size: 28 kDa
Immunohistochemistry staining of Calpain small subunit 1 in formalin-fixed, paraffin-embedded Human kidney tissue using 1/100 ab92333.
Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.
Overlay histogram showing HeLa cells stained with ab92333 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92333, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
ab92333 has been referenced in 4 publications.
- Li FZ et al. Crosstalk between calpain activation and TGF-ß1 augments collagen-I synthesis in pulmonary fibrosis. Biochim Biophys Acta 1852:1796-804 (2015). PubMed: 26071646
- Moore Z et al. NAMPT inhibition sensitizes pancreatic adenocarcinoma cells to tumor-selective, PAR-independent metabolic catastrophe and cell death induced by ß-lapachone. Cell Death Dis 6:e1599 (2015). PubMed: 25590809
- Cai JJ et al. Increased expression of Capn4 is associated with the malignancy of human glioma. CNS Neurosci Ther 20:521-7 (2014). PubMed: 24628706
- Guo Y et al. Remarkable Reduction of MAP2 in the Brains of Scrapie-Infected Rodents and Human Prion Disease Possibly Correlated with the Increase of Calpain. PLoS One 7:e30163 (2012). WB ; Chinese hamster . PubMed: 22272295