HSP70 Polyclonal antibody

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  WB analysis using 10995-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10995-1-AP (HSP70 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis using 10995-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 10995-1-AP (HSP70 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of human urine exosomes using 10995-1-AP

  human urine exosomes tissue were subjected to SDS PAGE followed by western blot with 10995-1-AP (HSP70 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IP experiment of mouse brain using 10995-1-AP

  IP result of anti-HSP70 (IP:10995-1-AP, 3ug; Detection:10995-1-AP 1:1000) with mouse brain tissue lysate 3600ug.

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  IHC staining of human breast cancer using 10995-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 10995-1-AP (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 colon using 10995-1-AP

  Immunohistochemical analysis of paraffin-embedded human normal colon slide using 10995-1-AP (HSP70 antibody) at dilution of 1:2000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human colon using 10995-1-AP

  Immunohistochemical analysis of paraffin-embedded human normal colon slide using 10995-1-AP (HSP70 antibody) at dilution of 1:2000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human liver cancer using 10995-1-AP

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 10995-1-AP (HSP70 Antibody) at dilution of 1:200 (under 40x lens).

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  IHC staining of mouse skin using 10995-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse skin tissue slide using 10995-1-AP (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 liver cancer using 10995-1-AP

  Immunohistochemical analysis of paraffin-embedded human liver cancer tissue slide using 10995-1-AP (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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  IF Staining of HeLa using 10995-1-AP

  Immunofluorescent analysis of (-20°C Methanol) fixed HeLa cells using HSP70 antibody (10995-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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  IF Staining of HepG2 using 10995-1-AP

  Immunofluorescent analysis of HepG2 cells, using HSPA1A antibody 10995-1-AP at 1:50 dilution and Rhodamine-labeled goat anti-rabbit IgG (red). Blue pseudocolor = DAPI (fluorescent DNA dye).

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  FC experiment of HeLa using 10995-1-AP

  1x10^6 HeLa cells were intracellularly stained with 0.25 ug HSP70 Polyclonal antibody (10995-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

• Featured Product
• KD/KO Validated

HSP70 antibodies are widely used to detect and study cellular stress responses by recognizing heat shock protein 70 in a variety of research applications, including cancer, neurodegeneration, and immune regulation. Proteintech has an extensive range of HSP70 antibodies validated for WB, IHC, IF/ICC, FC (Intra), IP, ELISA and more.

Cat No. 10995-1-AP

Publications(309)

Reviews (6)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (7)

Applications

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

Synonyms

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

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Tested Applications

• Positive WB detected in: A431 cells, human urine exosomes tissue, A549 cells, HeLa cells, Jurkat cells, mouse brain, HEK-293 cells, mouse kidney, rat kidney
• Positive IP detected in: mouse brain tissue
• Positive IHC detected in: human breast cancer tissue, human liver cancer tissue, human normal colon, mouse skin 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: HeLa cells, HepG2 cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:200-1:2000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 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

• KD/KO: See 9 publications below
• WB: See 283 publications below
• IHC: See 13 publications below
• IF: See 25 publications below
• IP: See 1 publications below
• ELISA: See 1 publications below
• FC: See 2 publications below
• CoIP: See 5 publications below

Product Information

10995-1-AP targets HSP70 in WB, IHC, IF/ICC, FC (Intra), IP, CoIP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, rabbit, chicken, zebrafish, sheep, goat, ostrich, oyster
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: HSP70 fusion protein Ag1446
• Full Name: heat shock 70kDa protein 1A
• Calculated Molecular Weight: 70 kDa
• Observed Molecular Weight: 66-70 kDa
• GenBank Accession Number: BC009322
• Gene Symbol: HSP70
• Gene ID (NCBI): 3303
• RRID: AB_2264230
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: P0DMV8
• 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

1. What is Hsp70/HSP1A?

   HSP1A is a member of the Hsp70 (heat shock protein 70) proteins that act as molecular chaperones ensuring correct protein folding and preventing protein aggregation. Hsp70 protein production is greatly induced by various stress stimuli, including high temperature and toxins. Its expression is often elevated in various cancers.

2. FAQs for Hsp70

   a. I cannot detect Hsp70 by western blotting

   HSP70s are typically expressed at low levels under normal physiological conditions but are dramatically up-regulated in response to cellular stress. Try to always include cell lysate from cells subjected to stress conditions as a positive control.

   b. What loading control can I use for cellular stress experiments with Hsp70?

   Choosing a loading control antibody is an important step in western blotting experimental setup. We highly recommend using more than one loading control while developing new cellular stress assays to ensure that a given treatment does not alter expression of house-keeping genes. Hsp70 has a molecular size of 70 kDa, so we recommend using GAPDH (36 kDa), actin (42 kDa), or tubulin (50-55 kDa). More information on our control antibodies can be found here: https://www.ptglab.com/news/blog/loading-control-antibodies-for-western-blotting/.

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
human	WB	Nature; A cytosolic surveillance mechanism activates the mitochondrial UPR; Authors - F X Reymond Sutandy
human	WB	Gastroenterology; PTEN deficiency facilitates exosome secretion and metastasis in cholangiocarcinoma by impairing TFEB-mediated lysosome biogenesis; Authors - Tian-Yi Jiang
human	WB	Nat Cell Biol; Caspase-2 is a condensate-mediated deubiquitinase in protein quality control; Authors - Yingwei Ge
human	WB	Nat Cell Biol; Heat stress activates YAP/TAZ to induce the heat shock transcriptome.; Authors - Min Luo ; KO Validated
human	WB	Mol Cancer; CircEZH2/miR-133b/IGF2BP2 aggravates colorectal cancer progression via enhancing the stability of m6A-modified CREB1 mRNA.; Authors - Bing Yao
mouse	IF	Adv Sci (Weinh); Reprogramming Lung Redox Homeostasis by NIR Driven Ultra-Small Pd Loaded Covalent Organic Framework Inhibits NF-κB Pathway for Acute Lung Injury Immunotherapy; Authors - Doudou Lei

Reviews

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

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FH

PK (Verified Customer) (08-14-2024)

• Applications: Western Blot
• Primary Antibody Dilution: 1000
• Cell Tissue Type: ac16

HSP70 Antibody Western Blot validation (1000 dilution) in ac16 (Cat no:10995-1-AP)

FH

Scott (Verified Customer) (05-22-2024)

20µg of protein was loaded and antibody was incubated overnight at 4oC following a total protein stain. The band appeared at the expected size. Precision plus protein standard ladder #1610373.

• Applications: Western Blot
• Primary Antibody Dilution: 1 : 5000
• Cell Tissue Type: TDP-Q331K Cortex and Spinal Cord Tissue Lysate

HSP70 Antibody Western Blot validation (1 : 5000 dilution) in TDP-Q331K Cortex and Spinal Cord Tissue Lysate (Cat no:10995-1-AP)

FH

Nin (Verified Customer) (01-28-2023)

A very good antibody to probe HSP70 by WB.

• Applications: Western Blot
• Primary Antibody Dilution: 1:2000
• Cell Tissue Type: mouse cortex lysate

HSP70 Antibody Western Blot validation (1:2000 dilution) in mouse cortex lysate (Cat no:10995-1-AP)

FH

Tsimafei (Verified Customer) (12-04-2022)

20µg of the total protein were used. Incubation was done for 2h at room temperature

• Applications: Western Blot
• Primary Antibody Dilution: 1:10000
• Cell Tissue Type: mESCs

HSP70 Antibody Western Blot validation (1:10000 dilution) in mESCs (Cat no:10995-1-AP)

FH

Guorong (Verified Customer) (03-22-2022)

Excellent performance with a specific band of expected size

• Applications: Western Blot
• Primary Antibody Dilution: 1:5000
• Cell Tissue Type: ARPE-19

HSP70 Antibody Western Blot validation (1:5000 dilution) in ARPE-19 (Cat no:10995-1-AP)

FH

Luxi (Verified Customer) (11-23-2020)

This antibody works well in my tissue. It binds to the nuclear and cytosol Hsp70s.

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:60
• Cell Tissue Type: embryonic cells