Anti-Calpain 1 抗体 [EPR3319]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Calpain 1 antibody [EPR3319] (AB108400)

ab108400 staining Calpain 1 in the T-47D cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/70). ab150077(1/500) an Alexa Fluor^®488-conjugated Goat anti-rabbit IgG was used as the secondary antibody. Nuclei were counterstained with DAPI.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 1 antibody [EPR3319] (AB108400)

ab108400 staining Calpain 1 in Human transitional cell carcinoma of bladder tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/200). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin. PBS was used in the negative control rather than the Primary antibody.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Calpain 1 antibody [EPR3319] (AB108400)

Overlay histogram showingJurkat cells stained with ab108400 (red line) at 1/380 dilution. The cells were fixed with 2% paraformaldehyde. The secondary antibody used was a FITC conjugated goat anti-rabbit IgG at 1/150 dilution. Isotype control antibody (green line) was rabbit monoclonal IgG used under the same conditions.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calpain 1 antibody [EPR3319] (AB108400)

ab108400, unpurified, at 1/500 dilution staining Calpain in Human kidney by Immunohistochemistry, Paraffin-embedded tissue.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Calpain 1 antibody [EPR3319] (AB108400)

ab108400, unpurified, at 1/250 dilution staining Calpain in HeLa cells by Immunofluorescence.

• WB

Lab

Western blot - Anti-Calpain 1 antibody [EPR3319] (AB108400)

ab108400 was shown to recognize Calpain 1 in wild-type HAP1. A faint cross-reactive band was observed in CAPN1 (Calpain 1) knockout cells at 82 kDa when over-exposed. Due to the immunogen's homology with CAPN2 (Calpain 2 Uniprot : P17655), this band could correspond to the Calpain 2 protein. Please note that cross-reactivity with this protein has not been confirmed experimentally.

All lanes:

Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400)

Lane 1:

Wild type HAP1 whole cell lysate

Lane 2:

CAPN1 (Calpain 1) knockout HAP1 whole cell lysate

Lane 3:

A431 whole cell lysate

Predicted band size: 82 kDa

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• WB

Supplier Data

Western blot - Anti-Calpain 1 antibody [EPR3319] (AB108400)

All lanes:

Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400) at 1/1000 dilution

All lanes:

C6 Lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), HRP- conjugated at 1/1000 dilution

Predicted band size: 62 kDa,82 kDa,87 kDa

Observed band size: 145 kDa,53 kDa

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• WB

Supplier Data

Western blot - Anti-Calpain 1 antibody [EPR3319] (AB108400)

All lanes:

Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400) at 1/5000 dilution

All lanes:

Human fetal kidney lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 82 kDa

false

• WB

Unknown

Western blot - Anti-Calpain 1 antibody [EPR3319] (AB108400)

All lanes:

Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400) at 1/1000 dilution

Lane 1:

Human fetal kidney cell lysate at 10 µg

Lane 2:

U937 cell lysate at 10 µg

Lane 3:

A431 cell lysate at 10 µg

Lane 4:

293T cell lysate at 10 µg

Lane 5:

T47D cell lysate at 10 µg

Predicted band size: 82 kDa

false

• WB

Supplier Data

Western blot - Anti-Calpain 1 antibody [EPR3319] (AB108400)

All lanes:

Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 20 µg

Lane 2:

Rat brain lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), HRP-conjugated at 1/1000 dilution

Predicted band size: 82 kDa

false

• WB

Supplier Data

Western blot - Anti-Calpain 1 antibody [EPR3319] (AB108400)

All lanes:

Western blot - Anti-Calpain 1 antibody [EPR3319] (ab108400) at 1/5000 dilution

All lanes:

T47-D Lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), HRP- conjugated at 1/1000 dilution

Predicted band size: 82 kDa

false

• WB

CiteAb

Western blot - Anti-Calpain 1 antibody [EPR3319] (AB108400)

Calpain 1 western blot using anti-Calpain 1 antibody [EPR3319] ab108400. Publication image and figure legend from Zeng, X., Chen, S., et al., 2017, Oncotarget, PubMed 29069832.

ab108400 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab108400 please see the product overview.

AG/UnAG attenuates calpain system activity in co-cultured myotubes(A) Western blot of μ-calpain, m-calpain, calpain-3, calpastatin and GAPDH in C2C12 myotubes. (B) Quantification of μ-calpain was normalized to GAPDH. Significant differences were detected between CO and any NC groups (#p < 0.001), between CO and CO+AG/UnAG groups (*p < 0.001), by one-way ANOVA followed by Tukey test. (C-E) Quantification of m-calpain, calpain-3 and calpastatin was normalized to GAPDH. No significant differences were detected among the six groups, by one-way ANOVA followed by Tukey test. (F) The levels of μ-calpain mRNA in C2C12 myotubes. mRNA levels were normalized to GAPDH. Significant differences were detected between NC and CO groups (#P = 0.005, P = 0.007; P = 0.007 respectively), between CO and CO+AG groups (*P = 0.01, P = 0.015 respectively), by one-way ANOVA followed by Tukey test. Data are represented as mean ± SD.

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