MA5-15578

antibody from Invitrogen Antibodies

Targeting: HSPA4 HS24/P52, HSPH2

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-15578

Provider

Invitrogen Antibodies

Product name

HSPA4 Monoclonal Antibody (5A6)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

MA5-15578 targets HSP70 in indirect ELISA, IF, IHC, and WB applications and shows reactivity with Human samples. The MA5-15578 immunogen is purified recombinant fragment of human HSP70 expressed in E. Coli. MA5-15578 detects HSP70 which has a predicted molecular weight of approximately 70kDa.

Reactivity

Human, Mouse

Host

Mouse

Isotype

IgG

Antibody clone number

5A6

Vial size

100 µL

Concentration

1.0 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Heteromeric complexes of heat shock protein 70 (HSP70) family members, including Hsp70B', in differentiated human neuronal cells.

Chow AM, Mok P, Xiao D, Khalouei S, Brown IR
Cell stress & chaperones 2010 Sep;15(5):545-53

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of HSPA4/Heat Shock 70 Protein 4 using HSPA4/Heat Shock 70 Protein 4 monoclonal antibody (Product # MA5-15578) in HeLa (1) cell lysate.

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), A-431 (Lane 2), HepG2 (Lane 3), MCF7 (Lane 4) and Mouse Testis (Lane 5). The blot was probed with Anti-HSPA4 Monoclonal Antibody (Product # MA5-15578, 1:2000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 1:4000 dilution). A 70kDa band corresponding to HSPA4 was observed across all cell lines and tissue lysates tested.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of NIH/3T3 cells using HSPA4/Heat Shock 70 Protein 4 monoclonal antibody (Product # MA5-15578) (Green). Blue: DRAQ5 fluorescent DNA dye.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of HSPA4 was performed using 70% confluent log phase HeLa cells. 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 HSPA4 Monoclonal Antibody (5A6) (Product # MA5-15578) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+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 ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing nucleus and cytoplasm localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded human breast cancer using HSPA4/Heat Shock 70 Protein 4 monoclonal antibody (Product # MA5-15578) followed with DAB staining.