HSP70 Monoclonal antibody

HSP70 Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat, yeast

Applications

WB, IHC, IF/ICC, FC (Intra), CoIP, ELISA

Conjugate

Unconjugated

Publications

23

CloneNo.

4E7E5

Reviews (1)

Cat no : 66183-1-Ig

Synonyms

HSPA1A, HSP72, HSP70.1, Heat shock protein family A member 1A, heat shock 70kDa protein 1A

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Validation Data Gallery

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  WB analysis using 66183-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66183-1-Ig (HSP70 antibody) at dilution of 1:50000 incubated at room temperature for 1.5 hours.

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  WB analysis of Recombinant protein using 66183-1-Ig

  6*His tagged-recombinant HSP70 (291-641 aa) and HSC70 (346-646 aa) proteins were subjected to SDS PAGE followed by western blot with 66183-1-Ig (HSP70 antibody), 66442-1-Ig (HSC70 antibody), and 66005-1-Ig (His antibody), respectively, at dilution of 1:50000, incubated at room temperature for 1.5 hours.

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  WB analysis of HEK-293 using 66183-1-Ig

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 66183-1-Ig (HSP70 Antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 66183-1-Ig

  HeLa cells were subjected to SDS PAGE followed by western blot with 66183-1-Ig (HSP70 Antibody) at dilution of 1:100000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HEK-293 using 66183-1-Ig

  HEK-293 cells were subjected to SDS PAGE followed by western blot with 66183-1-Ig (HSP70 Antibody) at dilution of 1:100000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of whole yeast using 66183-1-Ig

  whole yeast cells were subjected to SDS PAGE followed by western blot with 66183-1-Ig (HSP70 Antibody) at dilution of 1:100000 incubated at room temperature for 1.5 hours.

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  IHC staining of human breast cancer using 66183-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66183-1-Ig (HSP70 Antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 66183-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 66183-1-Ig (HSP70 Antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HEK-293 using 66183-1-Ig

  Immunofluorescent analysis of (-20℃ Ethanol ) fixed HEK-293 cells using 66183-1-Ig(HSP70 antibody) at dilution of 1:50 and Alexa Fluor 488-conjugated Goat Anti-Mouse IgG(H+L).

• 

  FC experiment of HeLa using 66183-1-Ig

  1X10^6 HeLa cells were intracellularly stained with 0.2 ug Anti-Human HSP70 (66183-1-Ig, Clone:4E7E5) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (green), and 0.2 ug Mouse IgG1 Isotype Control (66360-1-Ig, Clone: T1F8D3F10) (black). Cells were fixed with 4% PFA and permeabilized with 0.1% TritonX-100.

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Tested Applications

• Positive WB detected in: LNCaP cells, HEK-293 cells, HeLa cells, Recombinant protein, whole yeast, HEK-293T cells, K-562 cells
• Positive IHC detected in: human breast cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HEK-293 cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:20000-1:100000
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:20-1:200
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.20 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 18 publications below
• IHC: See 2 publications below
• IF: See 3 publications below
• ELISA: See 1 publications below
• CoIP: See 1 publications below

Product Information

66183-1-Ig targets HSP70 in WB, IHC, IF/ICC, FC (Intra), CoIP, ELISA applications and shows reactivity with human, mouse, rat, yeast samples.

• Tested Reactivity: human, mouse, rat, yeast
• Cited Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: HSP70 fusion protein Ag19111
• Full Name: heat shock 70kDa protein 1A
• Calculated Molecular Weight: 70 kDa
• Observed Molecular Weight: 65-70 kDa
• GenBank Accession Number: BC009322
• Gene Symbol: HSP70
• Gene ID (NCBI): 3303
• RRID: AB_2881578
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

HSPA1A , collectively known as HSP70 (also referred to HSP72), is a stress-inducible member of heat-shock protein 70 (HSP70) proteins which are highly conserved chaperons implicated in protein folding, protein refolding, protein transport, and protein targeting. Encoded by two closely linked, intronless and stress-inducible genes, HSPA1A and HSPA1B differ by only two amino acids and are believed to be fully interchangeable proteins. HSPA1A is a cytosol nuclear protein able to translocate between cytoplasm and nucleus. Generally, HSPA1A is thought to be expressed in unstressed normal cells at low or undetectable levels. Expression of HSPA1A protein can be highly activated by various stressful stimuli. Significant up-regulation of HSPA1A has been found in various tumors. Recently it has been reported that HSPA1A can be constitutively expressed in selected cell types. HSP70 is also used as exosomal marker. This antibody well recognized the endogenous HSP70 protein in multiple cell lines. (21373891)

Publications

Species	Application	Title
mouse	IF	Nat Commun; Autophagy-mediated apoptosis eliminates aneuploid cells in a mouse model of chromosome mosaicism.; Authors - Shruti Singla
rat	WB	Bioact Mater; Injectable cartilage matrix hydrogel loaded with cartilage endplate stem cells engineered to release exosomes for non-invasive treatment of intervertebral disc degeneration.; Authors - Liwen Luo
human	WB	Aging (Albany NY); MSC-derived exosomes promote recovery from traumatic brain injury via microglia/macrophages in rat.; Authors - Yunfei Chen
human	WB	Cells; Proteomics, Phosphoproteomics and Mirna Analysis of Circulating Extracellular Vesicles through Automated and High-Throughput Isolation.; Authors - Hao Zhang
human	WB	Mol Ther Oncolytics; Exosomal miR-205-5p enhances angiogenesis and nasopharyngeal carcinoma metastasis by targeting desmocollin-2.; Authors - Wenjuan Yang
mouse,human	IHC,CoIP	Genes (Basel); BAG3 Alleviates Atherosclerosis by Inhibiting Endothelial-to-Mesenchymal Transition via Autophagy Activation.; Authors - Hongtao Diao

Reviews

The reviews below have been submitted by verified Proteintech customers who received an incentive for providing their feedback.

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FH

Daniela (Verified Customer) (10-04-2022)

Very good specific antibody, no detection of no-specific bands in HeLa cell lysate. Two close bands at two different intensities are consistently detected below 75KDa in HeLa cell lysate.

• Applications: Western Blot, Immunoprecipitation
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: HeLa cells

HSP70 Antibody Western Blot,Immunoprecipitation validation (1:1000 dilution) in HeLa cells (Cat no:66183-1-Ig)