HSPA1B Polyclonal antibody

HSPA1B Polyclonal Antibody for WB, IHC, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF, IP, ELISA

Conjugate

Unconjugated

Publications

8

Cat no : 25405-1-AP

Synonyms

HSP70, HSP72

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Validation Data Gallery

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  WB analysis using 25405-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 25405-1-AP (HSP70 antibody) at dilution of 1:50000 incubated at room temperature for 1.5 hours.

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  WB analysis of rat brain using 25405-1-AP

  rat brain tissue were subjected to SDS PAGE followed by western blot with 25405-1-AP (HSP70 antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse brain using 25405-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 25405-1-AP (HSP70 antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of HEK-293 using 25405-1-AP

  IP result of anti-HSP70 (IP:25405-1-AP, 4ug; Detection:25405-1-AP 1:5000) with HEK-293 cells lysate 2320 ug.

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  IP experiment of K-562 using 25405-1-AP

  IP result of anti-HSP70 (IP:25405-1-AP, 4ug; Detection:25405-1-AP 1:1000) with K-562 cells lysate 3200ug.

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  IHC staining of human tonsillitis using 25405-1-AP

  Immunohistochemical analysis of paraffin-embedded human tonsillitis tissue slide using 25405-1-AP (HSP70 antibody) at dilution of 1:10000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human stomach using 25405-1-AP

  Immunohistochemical analysis of paraffin-embedded human stomach tissue slide using 25405-1-AP (HSPA1B antibody) at dilution of 1:10000 (under 20x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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Tested Applications

• Positive WB detected in: HEK-293 cells, mouse brain tissue, rat brain tissue, HeLa cells, K-562 cells
• Positive IP detected in: HEK-293 cells, K-562 cells
• Positive IHC detected in: human tonsillitis tissue, human stomach tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:50000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:5000-1:20000

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 7 publications below
• IF: See 1 publications below

Product Information

25405-1-AP targets HSPA1B in WB, IHC, IF, IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: HSPA1B fusion protein Ag13700
• Full Name: heat shock 70kDa protein 1B
• Calculated Molecular Weight: 70 kDa
• Observed Molecular Weight: 60-70 kDa
• GenBank Accession Number: BC057397
• Gene Symbol: HSP70
• Gene ID (NCBI): 3304
• RRID: AB_2880063
• 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. 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.

a. 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	Nat Commun; Feedback inhibition of cAMP effector signaling by a chaperone-assisted ubiquitin system.; Authors - Laura Rinaldi
mouse	WB	Hum Mol Genet; Loss of Ten-Eleven Translocation 2 induces cardiac hypertrophy and fibrosis through modulating ERK signaling pathway.; Authors - Huikang Tao
human	WB	Oncotarget; Quantitative proteomics reveals molecular mechanism of gamabufotalin and its potential inhibition on Hsp90 in lung cancer.; Authors - Liyuan Zhang
human	WB	Mol Cell Biol; An Important Role for RPRD1B in the Heat Shock Response; Authors - Simona Cugusi
human	WB	Front Biosci (Landmark Ed); Exosomal miR-22-3p from Mesenchymal Stem Cells Inhibits the Epithelial-Mesenchymal Transition (EMT) of Melanoma Cells by Regulating LGALS1; Authors - Yong Chen
human	WB	Cancer Gene Ther; Loss of exosomal miR-200b-3p from hypoxia cancer-associated fibroblasts promotes tumorigenesis and reduces sensitivity to 5-Flourouracil in colorectal cancer via upregulation of ZEB1 and E2F3; Authors - Wenjing Gong