MA3-014

antibody from Invitrogen Antibodies

Targeting: HSPA8 HSC70, HSC71, HSP73, HSPA10

Provider product page for MA3-014

Western blot

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [32]
Comments [0]
Validations
Immunocytochemistry [5]
Other assay [10]

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Product number: MA3-014 - Provider product page
Provider: Invitrogen Antibodies
Product name: HSC70 Monoclonal Antibody (13D3)
Antibody type: Monoclonal
Antigen: Other
Description: MA3-014 detects heat shock cognate protein 70 kDa (HSC70) from human, feline, primate, rat, and mouse tissues. This antibody displays slight cross-reactivity to HSP70. MA3-014 has been successfully used in Western blot, immunohistochemistry, immunofluorescence, and immunoprecipitation procedures By Western blot, this antibody detects a 70 kDa protein representing HSC70. By 2D gel electrophoresis, MA3-014 was found to bind strongly to HSC70 and faintly to HSP70 both before and after heat shock. Immunohistochemical staining of HSC70 in mouse spermatids/spermatozoa with MA3-014 results in staining restricted to the post acrosomal region in condensing spermatids and to the midpiece in spermatozoa. The MA3-014 antigen is mouse spermatogenic cell protein.
Reactivity: Human, Mouse, Rat, Feline
Host: Mouse
Isotype: IgM
Antibody clone number: 13D3
Vial size: 100 µL
Concentration: 1 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

HSPA8 protein structure - MA3-014 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of HSC70 using Monoclonal antibody (13D3) (Product # MA3-014) shows staining in NIH-3T3 cells. HSC70 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing HSC70 (Product # MA3-014) at a dilution of 1:20-1:200 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of HSC70 using HSC70 Monoclonal antibody (13D3) (Product # MA3-014) shows staining in HeLa cells. HSC70 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing HSC70 (Product # MA3-014) at a dilution of 1:20-1:200 over night at 4 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of HSC70 using HSC70 Monoclonal antibody (13D3) (Product # MA3-014) shows staining in MCF-7 cells. HSC70 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing HSC70 (Product # MA3-014) at a dilution of 1:20-1:200 °C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of HSC70/Heat Shock Cognate Protein was performed using 70% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HSC70/Heat Shock Cognate Protein Mouse Monoclonal Antibody (Product # MA3-014) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L)/IgM (L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Heat Shock Protein 70 (HSC70) (green) in HeLa and NIH3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a HSC70 monoclonal antibody (Product # MA3-014) at a dilution of 1:50 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunoprecipitation of HSC70 was performed on HeLa cells. Antigen-antibody complexes were formed by incubating 500 µg whole cell lysate with 2 µL of HSC70 monoclonal antibody (Product # MA3-014) overnight on a rocking platform at 4°C. The immune complexes were captured on 50 µL Protein A/G Plus Agarose (Product # 20421), washed extensively, and eluted with Lane Marker Reducing Sample Buffer (Product # 39000). Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a HSC70 monoclonal antibody (Product # MA3-014) at a dilution of 1:1000 overnight rotating at 4°C, washed in TBST, and probed with goat anti-mouse IgM-HRP secondary antibody (Product # 31440) for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Validation of HSC70 immunohistochemical staining. (a, b) HSC70 knockdown in NB4 APL cells. (a) HSC70 knockdown efficiency of three independent shRNAs (shHSC70_1-3) was determined by western blotting and comparison to scramble shRNA transduced control cells (SHC002). (b) The most efficient HSC70 knockdown (shHSC70_3) was selected and subjected to immunohistochemistry. (c, d) 293 T cells were transiently transfected with an empty vector (ctrl) and HSC70 expression plasmid. (c) HSC70 expression was validated by western blotting. (d) 293 T cells were subjected to HSC70 immunohistochemistry.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 HSPA8, examples of immunohistochemical staining: ( A ): Adenocarcinoma, IRS 3 x 3 = 9; ( B ) Adenocarcinoma, IRS 2 x 4 = 8; ( C ): Squamous cell carcinoma, IRS 2 x 4 = 8; ( D ): Squamous cell carcinoma, IRS 0 x 4 = 0; Objective magnification: 10x, Scale bar: 200 um.