Recombinant Anti-Hsp70 antibody [3A3] (ab5439)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [3A3] to Hsp70
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-Hsp70 antibody [3A3]
See all Hsp70 primary antibodies -
Description
Mouse monoclonal [3A3] to Hsp70 -
Host species
Mouse -
Specificity
This antibody detects several members of the heat shock protein 70 kDa (Hsp 70) gene family including Hsp 70, Hsc 70, p75, and following heat shock, Hsp 72 from yeast, Drosophila, fish, mouse, avian, amphibian and human samples.
-
Tested applications
Suitable for: ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment corresponding to Human Hsp70 aa 504-617 (C terminal).
-
Epitope
Epitope mapping with a panel of Hsp 70 deletion mutants suggests that the epitope recognized is located between amino acids 504-617 of human Hsp 70, a region that has been shown to be involved in stress-induced nucleolar localization. -
Positive control
- WB: HeLa, K562, Jurkat, 293T, A431, A549, F9, RAW264.7 and C6, PC-12 lysates. IHC-P: Human cerebrum and lung carcinoma, Mouse testis, Rat cerebrum tissues. ICC/IF: HeLa and NIH/3T3, PC-12 cells.
-
General notes
This product has switched from a hybridoma to recombinant production method on 08th March 2021.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
Purity
Protein A purified -
Primary antibody notes
The Hsp 70 family is a set of highly conserved proteins that are induced by a variety of biological stresses, including heat stress, in every organism in which the proteins have been examined. The human Hsp 70 family members include: Hsp 70, a protein which is strongly inducible in all organisms but which is also constitutively expressed in primate cells; Hsp 72, a 72 kDa protein that is induced exclusively under stress conditions; Hsc 70, or cognate protein, is a 72 kDa, constitutively expressed, protein which is involved in the uncoating of clathrin coated vesicles; GRP78, or BiP, is a glucose regulated 78 kDa protein localized in the endoplasmic reticulum; and p75, or Hsp 75, a 75 kDa protein that is found within the mitochondria. -
Clonality
Monoclonal -
Clone number
3A3 -
Isotype
IgG1 -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab5439 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
ICC/IF |
1/50.
|
|
IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
WB | (3) |
1/1000. Predicted molecular weight: 70-78 kDa.
|
Notes |
---|
ICC/IF
1/50. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Predicted molecular weight: 70-78 kDa. |
Target
-
Relevance
Function: In cooperation with other chaperones, the Hsp70 family stabilize preexistent proteins against aggregation and mediate the folding of newly translated polypeptides in the cytosol as well as within organelles. These chaperones participate in all these processes through their ability to recognize nonnative conformations of other proteins. They bind extended peptide segments with a net hydrophobic character exposed by polypeptides during translation and membrane translocation, or following stress-induced damage. In case of rotavirus A infection, serves as a post-attachment receptor for the virus to facilitate entry into the cell. Tissue specificity: HSPA1B is testis-specific. -
Cellular localization
Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. -
Database links
- Entrez Gene: 15511 Human
- Entrez Gene: 3303 Human
- Entrez Gene: 193740 Mouse
- Entrez Gene: 3304 Mouse
- Entrez Gene: 24472 Rat
- Entrez Gene: 294254 Rat
- Omim: 140550 Human
- Omim: 603012 Human
see all -
Alternative names
- DnaK type molecular chaperone HSP70 1 antibody
- Epididymis secretory protein Li 103 antibody
- FLJ54303 antibody
see all
Images
-
All lanes : Anti-Hsp70 antibody [3A3] (ab5439) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lane 4 : 293T (human embryonic kidney epithelial cell), whole cell lysate
Lane 5 : A431 (human epidermoid carcinoma epithelial cell), whole cell lysate
Lane 6 : A549 (human lung carcinoma epithelial cell), whole cell lysate
Lane 7 : F9 (mouse embryonal carcinoma epithelial cell), whole cell lysate
Lane 8 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lane 9 : C6 (rat glial tumor glial cell), whole cell lysate
Lane 10 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Predicted band size: 70-78 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking/Diluting buffer and concentration: 5% NFDM/TBST
-
Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Hsp70 with ab5439 at 1/2000 (0.519 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum. The section was incubated with ab5439 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling Hsp70 with ab5439 at 1/50 (20.76 µg/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and weak nuclear staining in HeLa cells ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
-
Immunohistochemical analysis of paraffin-embedded Human lung carcinoma tissue labeling Hsp70 with ab5439 at 1/2000 (0.519 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human lung carcinoma. The section was incubated with ab5439 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling Hsp70 with ab5439 at 1/50 (20.76 µg/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and weak nuclear staining in NIH/3T3 cells ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
-
Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Hsp70 with ab5439 at 1/2000 (0.519 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse testis. The section was incubated with ab5439 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
-
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-12 cells labelling Hsp70 with ab5439 at 1/50 (20.76 µg/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in PC-12 cells ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
-
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling Hsp70 with ab5439 at 1/2000 (0.519 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat cerebrum. The section was incubated with ab5439 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (57)
ab5439 has been referenced in 57 publications.
- Mohan HM et al. RTL8 promotes nuclear localization of UBQLN2 to subnuclear compartments associated with protein quality control. Cell Mol Life Sci 79:176 (2022). PubMed: 35247097
- Hu C et al. Heat Stress-Induced Dysbiosis of Porcine Colon Microbiota Plays a Role in Intestinal Damage: A Fecal Microbiota Profile. Front Vet Sci 9:686902 (2022). PubMed: 35300220
- Zhu J et al. Extracellular Vesicle-Derived circITGB1 Regulates Dendritic Cell Maturation and Cardiac Inflammation via miR-342-3p/NFAM1. Oxid Med Cell Longev 2022:8392313 (2022). PubMed: 35615580
- Mastroiacovo F et al. Within the Ischemic Penumbra, Sub-Cellular Compartmentalization of Heat Shock Protein 70 Overlaps with Autophagy Proteins and Fails to Merge with Lysosomes. Molecules 27:N/A (2022). PubMed: 35630599
- Gómez-Marín E et al. The high mobility group protein HMG20A cooperates with the histone reader PHF14 to modulate TGFβ and Hippo pathways. Nucleic Acids Res 50:9838-9857 (2022). PubMed: 36124662