Rabbit anti-Human HSP70 Monoclonal Antibody | anti-HSPA1A antibody

Recombinant Anti-HSP70 Antibody, Rabbit Monoclonal

Cat. Num. AAA8105583

• Gene Names: HSPA1B; HSP72; HSPA1; HSX70; HSP70-1; HSP70-2; HSP70.1; HSP70.2; HSP70-1B
• Reactivity: Human
• Applications: Western Blot, ELISA, Immunohistochemistry, Flow Cytometry, Functional Assay, Immunocytochemistry, Immunofluorescence, Immunoprecipitation
• Purity: Protein A
• Synonyms: HSP70; Monoclonal Antibody; Recombinant Anti-HSP70 Antibody; Rabbit Monoclonal; HSP70/HSPA1A Antibody; Rabbit MAb; heat shock 70kDa protein 1A; Anti-HEL-S-103 Antibody; Anti-HSP70-1 Antibody; Anti-HSP70-1A Antibody; Anti-HSP70I Antibody; Anti-HSP72 Antibody; Anti-HSPA1 Antibody; anti-HSPA1A antibody

• Host: Rabbit
• Reactivity: Human
• Clonality: Monoclonal
• Isotype: Rabbit IgG
• Clone Number: 001
• Specificity: Human HSP70/HSPA1A
• Purity/Purification: Protein A
• Form/Format: Liquid; 0.2um filtered solution in PBS

• Applicable Applications for anti-HSPA1A antibody: Western Blot (WB), ELISA (EIA), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunoprecipitation (IP)
• Application Notes: WB: 1:500-1:2000; ELISA: 1:5000-1:10,000; IHC-P: 1:500-1:2000; ICC/IF: 1:20-1:100; FC/FACS/FCM: 1:25-1:100; IP: 1-4uL/mg of lysate
• Antibody Type: Recombinant Antibody
• Immunogen: Recombinant Human HSP70/HSPA1A protein
• Conjugation: Unconjugated
• Preparation: This antibody was obtained from a rabbit immunized with purified, recombinant Human HSP70/HSPA1A (rh HSP70/HSPA1A).
• Preparation and Storage: This antibody can be stored at 2-8 degree C for one month without detectable loss of activity. Antibody products are stable for twelve months from date of receipt when stored at -20 degree C to -80 degree C. Preservative-Free. Avoid repeated freeze-thaw cycles.

Immunofluorescence (IF)

(Immunofluorescence staining of Human HSPA1A in Hela cells. Cells were fixed with 4% PFA, permeabilzed with 0.3% Triton X-100 in PBS, blocked with 10% serum, and incubated with rabbit anti-Human HSPA1A monoclonal antibody (1:60) at 37 degree C 1 hour. Then cells were stained with the Alexa Fluor 594-conjugated goat Anti-rabbit IgG secondary antibody (red) and counterstained with DAPI (blue). Positive staining was localized to cytoplasm.)

Immunohistochemistry (IHC)

(Immunochemical staining of HSPA1A in human hepatoma with rabbit monoclonal antibody (1:1000, formalin-fixed paraffin embedded sections).)

Immunohistochemistry (IHC)

(Immunochemical staining of HSPA1A in cynomolgus macaque testis with rabbit monoclonal antibody (1:1000, formalin-fixed paraffin embedded sections).)

Flow Cytometry (FC/FACS)

(Flow cytometric analysis of Human HSPA1A expression on HeLa cells. The cells were treated according to manufacturer's manual (BD Pharmingen'), stained with purified anti-Human HSPA1A, then a FITC-conjugated second step antibody. The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells.)

Western Blot (WB)

(Anti-HSPA1A rabbit monoclonal antibody at 1:500 dilution Lane A: A549 whole cell lysate Lane B: HCT116 whole cell lysate Lane C: Hela whole cell lysate Lane D: HepG2 whole cell lysate Lane E: HL60 whole cell lysate Lane F: A431 whole cell lysate Lane G: MCF7 whole cell lysate Lane H: K562 whole cell lysate Lane I: C6 whole cell lysate Lane J: 293T whole cell lysate Lane K: MOLT4 whole cell lysate Lane L: Jurkat whole cell lysate Lysates/proteins at 30 ug per lane. Secondary Goat Anti-Rabbit IgG H&L (Dylight800) at 1/10000 dilution. Developed using the Odyssey technique. Performed under reducing conditions. Predicted band size:70 kDa Observed band size:70 kDa (We are unsure as to the identity of these extra bands.))

Immunoprecipitation (IP)

(HSPA1A was immunoprecipitated using:Lane A:0.5 mg Hela Whole Cell Lysate2 uL anti-HSPA1A rabbit monoclonal antibody and 15 ul of 50 % Protein G agarose.Primary antibody:Anti-HSPA1A rabbit monoclonal antibody,at 1:200 dilution Secondary antibody:Dylight 800-labeled antibody to rabbit IgG (H+L), at 1:5000 dilution Developed using the odssey technique.Performed under reducing conditions.Predicted band size: 70 kDaObserved band size: 70 kDa)

Related Product Information for anti-HSPA1A antibody

HSPA1A is a member of the Hsp70 protein family. The 70 kilodalton heat shock proteins (Hsp70s) are a family of ubiquitously expressed heat shock proteins. HSP are abundant and conserved proteins present in all cells. Upon temperature shock or other stress stimuli, HSP is synthesized intracellularly, which may protect cells from protein denaturation or death. Extracellularly, HSP can serve a cytokine function to initiate both innate and adaptive immunity through activation of APC. HSP serves also a chaperone function and facilitates the presentation of antigen peptide to T cells. Molecular chaperones of the Hsp70 family have diverse functions in cells. They assist the folding of newly synthesized and stress-denatured proteins, as well as the import of proteins into organelles, and the dissociation of aggregated proteins. The well-conserved Hsp70 chaperones are ATP dependent: binding and hydrolysis of ATP regulate their interactions with unfolded polypeptide substrates, and ATPase cycling is necessary for their function. All cellular functions of Hsp70 chaperones use the same mechanism of ATP-driven polypeptide binding and release.

Product Categories/Family for anti-HSPA1A antibody

Death Receptor Signaling

References

Heck TG, et al. (2011) HSP70 expression: does it a novel fatigue signalling factor from immune system to the brain Cell Biochem Funct. 29 (3): 215-26.Chen T, et al. (2010) Stress for maintaining memory: HSP70 as a mobile messenger for innate and adaptive immunity. Eur J Immunol. 40 (6): 1541-4.Young JC. (2010) Mechanisms of the Hsp70 chaperone system. Biochem Cell Biol. 88 (2): 291-300.

NCBI and Uniprot Product Information

• NCBI GI #: 167466173
• NCBI GeneID: 3304
• NCBI Accession #: NP_005337.2
• NCBI GenBank Nucleotide #: NM_005346.4
• UniProt Accession #: P0DMV9; P08107; P19790; Q5JQI4; Q5SP17; Q9UQL9; Q9UQM0; B4E3B6
• Molecular Weight: 70,052 Da
• NCBI Official Full Name: heat shock 70 kDa protein 1B
• NCBI Official Synonym Full Names: heat shock protein family A (Hsp70) member 1B
• NCBI Official Symbol: HSPA1B
• NCBI Official Synonym Symbols: HSP72; HSPA1; HSX70; HSP70-1; HSP70-2; HSP70.1; HSP70.2; HSP70-1B
• NCBI Protein Information: heat shock 70 kDa protein 1B
• UniProt Protein Name: Heat shock 70 kDa protein 1B
• Protein Family: HSP70 co-chaperone
• UniProt Gene Name: HSPA1B
• UniProt Synonym Gene Names: HSP721 PublicationManual assertion based on opinion iniRef.36; HSP70-22 PublicationsManual assertion based on opinion iniRef.3; HSP70.2

NCBI Description

This intronless gene encodes a 70kDa heat shock protein which is a member of the heat shock protein 70 family. In conjuction with other heat shock proteins, this protein stabilizes existing proteins against aggregation and mediates the folding of newly translated proteins in the cytosol and in organelles. It is also involved in the ubiquitin-proteasome pathway through interaction with the AU-rich element RNA-binding protein 1. The gene is located in the major histocompatibility complex class III region, in a cluster with two closely related genes which encode similar proteins. [provided by RefSeq, Jul 2008]

Uniprot Description

Molecular chaperone implicated in a wide variety of cellular processes, including protection of the proteome from stress, folding and transport of newly synthesized polypeptides, activation of proteolysis of misfolded proteins and the formation and dissociation of protein complexes. Plays a pivotal role in the protein quality control system, ensuring the correct folding of proteins, the re-folding of misfolded proteins and controlling the targeting of proteins for subsequent degradation. This is achieved through cycles of ATP binding, ATP hydrolysis and ADP release, mediated by co-chaperones. The co-chaperones have been shown to not only regulate different steps of the ATPase cycle, but they also have an individual specificity such that one co-chaperone may promote folding of a substrate while another may promote degradation. The affinity for polypeptides is regulated by its nucleotide bound state. In the ATP-bound form, it has a low affinity for substrate proteins. However, upon hydrolysis of the ATP to ADP, it undergoes a conformational change that increases its affinity for substrate proteins. It goes through repeated cycles of ATP hydrolysis and nucleotide exchange, which permits cycles of substrate binding and release. The co-chaperones are of three types: J-domain co-chaperones such as HSP40s (stimulate ATPase hydrolysis by HSP70), the nucleotide exchange factors (NEF) such as BAG1/2/3 (facilitate conversion of HSP70 from the ADP-bound to the ATP-bound state thereby promoting substrate release), and the TPR domain chaperones such as HOPX and STUB1 (PubMed:24012426, PubMed:26865365, PubMed:24318877). Maintains protein homeostasis during cellular stress through two opposing mechanisms: protein refolding and degradation. Its acetylation/deacetylation state determines whether it functions in protein refolding or protein degradation by controlling the competitive binding of co-chaperones HOPX and STUB1. During the early stress response, the acetylated form binds to HOPX which assists in chaperone-mediated protein refolding, thereafter, it is deacetylated and binds to ubiquitin ligase STUB1 that promotes ubiquitin-mediated protein degradation (PubMed:27708256). Regulates centrosome integrity during mitosis, and is required for the maintenance of a functional mitotic centrosome that supports the assembly of a bipolar mitotic spindle (PubMed:27137183). Enhances STUB1-mediated SMAD3 ubiquitination and degradation and facilitates STUB1-mediated inhibition of TGF-beta signaling (PubMed:24613385). Essential for STUB1-mediated ubiquitination and degradation of FOXP3 in regulatory T-cells (Treg) during inflammation (PubMed:23973223).