Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunohistochemistry [3]
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Validation data
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- Product number
- MA1-13079 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Calpain 1 Monoclonal Antibody (P-6(2H7))
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- MA1-13079 detects calpain-1 large subunit (mu-type) in human and mouse samples by western blot, immunofluorescence, immunohistochemistry.
- Reactivity
- Human, Mouse
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- P-6(2H7)
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Calpain-1 was performed by loading 20µg of indicated whole cell lysates and 7µl of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BX10). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% Milk in TBST for 1 hour at room temperature. Calpain-1 was detected at ~80kDa using a Calpain1 mouse monoclonal antibody (Product # MA1-13079) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform followed by a Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177) at a dilution of 1:2000 for at least 30 minutes at room temperature. GAPDH was detected using a GAPDH rabbit polyclonal antibody, HRP conjugate (Product # PA1-987-HRP) at a dilution of 1:1000 overnight at 4C on a rocking platform. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- MA1-13079 Calpain-1 P-6 (2H7) Antibody in WB-1 Western blot analysis of Calpain-1 was performed by loading 20µg of indicated whole cell lysates and 7 µL of PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel (Product # WT4202BX10). Proteins were transferred to a nitrocellulose membrane using the G2 Blotter (Product # 62288), and blocked with 5% Milk in TBST for 1 hour at room temperature. Calpain-1 was detected at ~80 kDa using a Calpain1 mouse monoclonal antibody (Product # MA1-13079) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform followed by a Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate at a dilution of 1:1000 for at least 30 minutes at room temperature. GAPDH was detected using a GAPDH rabbit polyclonal antibody, HRP conjugate (Product # PA1-987-HRP) at a dilution of 1:1000 overnight at 4C on a rocking platform. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on human colon carcinoma tissue. Tissue was deparaffinized with xylene, followed by rehydration in sequential washes of 100% ethanol, 95% ethanol, 80% ethanol, and water. To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0) and heated for 20 min. Following antigen retrieval, tissues were blocked in a 3% BSA in wash buffer solution for 30-60 minutes at room temperature. Tissue was then probed with a Calpain-1 Mouse Monoclonal antibody (Product # MA1-13079) at a dilution of 1:100 in blocking buffer overnight at 4°C in a humidified chamber (right panel). Negative control tissue received no primary antibody (right panel). Tissues were washed extensively with PBST, and detection was performed using a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:500 followed by colorimetric detection using metal enhanced DAB. Tissues were then counterstained with hematoxylin and prepped for mounting and imaging.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on human human kidney tissue. Tissue was deparaffinized with xylene, followed by rehydration in sequential washes of 100% ethanol, 95% ethanol, 80% ethanol, and water. To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0) and heated for 20 min. Following antigen retrieval, tissues were blocked in a 3% BSA in wash buffer solution for 30-60 minutes at room temperature. Tissue was then probed with a Calpain-1 mouse monoclonal antibody (Product # MA1-13079) at a dilution of 1:100 in blocking buffer overnight at 4°C in a humidified chamber (right panel). Negative control tissue received no primary antibody (left panel). Tissues were washed extensively with PBST, and detection was performed using a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:500 followed by colorimetric detection using metal enhanced DAB. Tissues were then counterstained with hematoxylin and prepped for mounting and imaging.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry was performed on mouse colon tissue. Tissue was deparaffinized with xylene, followed by rehydration in sequential washes of 100% ethanol, 95% ethanol, 80% ethanol, and water. To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0) and heated for 20 minutes. Following antigen retrieval, tissues were blocked in a 3% BSA in wash buffer solution for 30-60 minutes at room temperature. Tissue was then probed with a Calpain-1 mouse monoclonal antibody (Product # MA1-13079) at a dilution of 1:20 in blocking buffer overnight at 4°C in a humidified chamber (left panel). Negative control tissue received no primary antibody (right panel). Tissues were washed extensively with PBST, and detection was performed using a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:500 followed by colorimetric detection using metal enhanced DAB. Tissues were then counterstained with hematoxylin and prepped for mounting and imaging.