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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [2]
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- Validations
- Western blot [2]
- Immunocytochemistry [1]
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- Product number
- MA1-26078 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- HSC70 Monoclonal Antibody (1B5)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- Recommended positive controls: Bovine Hsc70 Protein and HeLa Heat Shocked Cell Lysate.
- Antibody clone number
- 1B5
- Concentration
- 1 mg/mL
Submitted references Proteasome inhibition alleviates SNARE-dependent neurodegeneration.
CSPα promotes SNARE-complex assembly by chaperoning SNAP-25 during synaptic activity.
Sharma M, Burré J, Südhof TC
Science translational medicine 2012 Aug 15;4(147):147ra113
Science translational medicine 2012 Aug 15;4(147):147ra113
CSPα promotes SNARE-complex assembly by chaperoning SNAP-25 during synaptic activity.
Sharma M, Burré J, Südhof TC
Nature cell biology 2011 Jan;13(1):30-9
Nature cell biology 2011 Jan;13(1):30-9
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), PC3 (Lane 2), CaCO-2 (Lane 3), Hep G2 (Lane 4), K-562 (Lane 5), Raji (Lane 6), SH-SY5Y (Lane 7), HL-60 (Lane 8) and A431 (Lane 9). The blot was probed with Anti-HSC70 Monoclonal Antibody (1B5) (Product # MA1-26078, 0.5 µg/ml) and detected by chemiluminescence using F(ab')2-Rabbit anti-Rat IgG (H+L) Secondary Antibody, HRP (Product # PA1-29927, 0.25 µg/ml, 1:4000 dilution). A 70 kDa band corresponding to HSC70 was observed across all the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of HSC70 was achieved by transfecting HeLa with HSC70 specific siRNAs (Silencer® select Product # s6987). Western blot analysis (Fig. a) was performed using whole cell extracts from the HSC70 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with HSC70 Monoclonal Antibody (Product # MA1-26078, 1µg/ml) and F(ab')2- Rabbit anti-Rat IgG (H+L) Secondary Antibody, HRP conjugate (Product # PA1-29927, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to HSC70.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of HSC70 was performed using 70% confluent log phase PC-3 cells treated with Heat Shock (43 degree celsius for 1 hr). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HSC70 Monoclonal Antibody (1B5) (Product # MA1-26078) at 1 µg/mL concentration in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Product # A-11006) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing HSC70 in the nucleolus and cytoplasm. Panel e represents the untreated cells showing cytoplasmic and faint nucleus localization. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
