HSP70 Polyclonal antibody

• Featured Product
• KD/KO Validated

HSP70 Polyclonal Antibody for FC, IF, IHC, IP, WB, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat and More (5)

Applications

WB, IP, IHC, IF, FC, CoIP, ELISA

Conjugate

Unconjugated

Publications

154

Reviews (3)

Cat no : 10995-1-AP

Synonyms

Heat shock 70 kDa protein 1/2, heat shock 70kDa protein 1A, HSP70, HSP70 1, HSP70 1/HSP70 2, HSP70 1A, HSP70.1/HSP70.2, HSP70I, HSP72, HSPA1, HSPA1A, HSPA1B

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Validation Data Gallery

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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.

• 

  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.

• 

  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.

• 

  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).

• 

  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).

• 

  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).

• 

  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).

• 

  IF Staining of HeLa using 10995-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using 10995-1-AP (HSP70 antibody), at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L). F-actin was stained with CL594-phalloidin.

• 

  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).

• 

  FC experiment of HepG2 using 10995-1-AP

  1X10^6 HepG2 cells were stained with 0.2ug HSP70 antibody (10995-1-AP, red) and control antibody (blue). Fixed with 90% MeOH blocked with 3% BSA (30 min). Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) with dilution 1:1000.

• 

  FC experiment of HeLa using 10995-1-AP

  1X10^6 HeLa cells were stained with 0.20ug HSP70 antibody (10995-1-AP, red) and control antibody (blue). Fixed with 90% MeOH.

• 

  FC experiment of A549 using 10995-1-AP

  1X10^6 A549 cells were stained with 0.20ug HSP70 antibody (10995-1-AP, red) and control antibody (blue). Fixed with 90% MeOH.

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Tested Applications

• Positive WB detected in: A431 cells, HEK-293 cells, HeLa cells, Jurkat cells, mouse kidney, rat kidney, A549 cells, mouse brain
• Positive IP detected in: mouse brain tissue
• Positive IHC detected in: human breast cancer tissue, human liver cancer tissue, 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 detected in: HeLa cells, HepG2 cells
• Positive FC detected in: HepG2 cells, HeLa cells, A549 cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for IP and 1:500-1:2000 for WB
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF): IF : 1:50-1:500

Sample-dependent, check data in validation data gallery

Published Applications

• KD/KO: See 7 publications below
• WB: See 145 publications below
• IHC: See 7 publications below
• IF: See 14 publications below
• IP: See 1 publications below
• FC: See 1 publications below
• CoIP: See 2 publications below

Product Information

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

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse, rat, bovine, C. elegans, ostrich, oyster, pig
• 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: HSPA1A
• Gene ID (NCBI): 3303
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity 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.

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

Species	Application	Title
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; Heat stress activates YAP/TAZ to induce the heat shock transcriptome.; Authors - Min Luo ; KO Validated
human,mouse	WB,IF	Acta Neuropathol; Distinctive chaperonopathy in skeletal muscle associated with the dominant variant in DNAJB4; Authors - Michio Inoue
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	WB	Neuron; UBQLN2-HSP70 axis reduces poly-Gly-Ala aggregates and alleviates behavioral defects in the C9ORF72 animal model.; Authors - Kejing Zhang ; KD Validated
human,mouse	WB,IHC	Sci Adv; Single-cell analysis of human basal cell carcinoma reveals novel regulators of tumor growth and the tumor microenvironment.; Authors - Christian F Guerrero-Juarez

Reviews

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

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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

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