Clone:
REA349
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Alternative names:
HSPA1B, HSPA1A, HSP70-1, HSP70-2, HSP70.1, HSP70.2

Extended validation for Hsp70 Antibody, anti-human/mouse, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA126
W27++
Cells were incubated with an excess of purified unconjugated Hsp70 (REA349) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for Hsp70. HeLa cells were first stained with Viobility™ Fixable Dye. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with Hsp70 antibodies and plotted against the side scatter. As a control, Hsp70 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for Hsp70. HeLa cells were first stained with Viobility™ Fixable Dye. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with Hsp70 antibodies and plotted against the side scatter. As a control, Hsp70 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for Hsp70. HeLa cells were first stained with Viobility™ Fixable Dye. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with Hsp70 antibodies and plotted against the side scatter. As a control, Hsp70 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for Hsp70. HeLa cells were first stained with Viobility™ Fixable Dye. Cells were then fixed and permeabilized using the Inside Stain Kit followed by a staining with Hsp70 antibodies and plotted against the side scatter. As a control, Hsp70 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for Hsp70 Antibody, anti-human/mouse, REAfinity™

Overview

Clone REA349 recognizes the human and mouse heat shock 70 kDa protein (Hsp70) antigen, a member of the heat shock protein family that serve critical roles in protein homeostasis. The Hsp70 family is the most conserved in evolution, and all eukaryotes contain multiple members. Their synthesis is increased upon environmental stress. Hsp70 assists a wide range of folding processes, including the folding and assembly of newly synthesized proteins, refolding of misfolded and aggregated proteins, membrane translocation of organellar and secretory proteins, and control of the activity of regulatory proteins. It has thus housekeeping functions in the cell in which it is built-in components of folding and signal transduction pathways, and quality control functions in which it proofreads the structure of proteins and repairs misfolded conformers. All of these activities appear to be based on the property of Hsp70 to interact with hydrophobic peptide segments of proteins in an ATP-controlled fashion.
Additional information: Clone REA349 displays negligible binding to Fc receptors.

Alternative names

HSPA1B, HSPA1A, HSP70-1, HSP70-2, HSP70.1, HSP70.2

Detailed product information

Technical specifications

CloneREA349
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, mouse
AntigenHsp70
Alternative names of antigenHSPA1B, HSPA1A, HSP70-1, HSP70-2, HSP70.1, HSP70.2
Molecular mass of antigen [kDa]70(refers to membrane-bound form)
Entrez Gene ID3303, 3304
RRIDAB_2857706, AB_2857423, AB_2857421, AB_2857829, AB_2857822, AB_2857745, AB_2857739, AB_2857708

Resources for Hsp70 Antibody, anti-human/mouse, REAfinity™

Certificates

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References for Hsp70 Antibody, anti-human/mouse, REAfinity™

Publications

  1. Murphy, M. E. et al. (2013) The HSP70 family and cancer. Carcinogenesis 34(6): 1181-1188
  2. Milner, C. M. et al. (1990) Structure and expression of the three MHC-linked HSP70 genes. Immunogenetics 32(4): 242-251
  3. Mayer, M. P. et al. (2005) Hsp70 chaperones: cellular functions and molecular mechanism. Cell. Mol. Life Sci. 62(6): 670-684

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