HSPA4 Polyclonal antibody

• Featured Product
• KD/KO Validated

HSPA4 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat, monkey

Applications

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

Conjugate

Unconjugated

Publications

4

Cat no : 21206-1-AP

Synonyms

HSPH2, hsp70RY, Heat shock protein family H member 2, Heat shock 70-related protein APG-2, Heat shock 70 kDa protein 4

---

Validation Data Gallery

• 

  WB analysis using 21206-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 21206-1-AP (HSPA4 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HEK-293 using 21206-1-AP

  WB result of HSPA4 antibody (21206-1-AP; 1:2000; incubated at room temperature for 1.5 hours) with sh-Control and sh-HSPA4 transfected HEK-293 cells.

• 

  WB analysis of NIH/3T3 using 21206-1-AP

  NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 21206-1-AP (HSPA4 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse testis using 21206-1-AP

  mouse testis tissue were subjected to SDS PAGE followed by western blot with 21206-1-AP (HSPA4 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of mouse brain using 21206-1-AP

  IP result of anti-HSPA4 (IP:21206-1-AP, 4ug; Detection:21206-1-AP 1:1000) with mouse brain tissue lysate 4000ug.

• 

  IHC staining of human colon cancer using 21206-1-AP

  Immunohistochemical analysis of paraffin-embedded human colon cancer using 21206-1-AP (HSPA4 antibody) at dilution of 1:100 (under 10x lens).

• 

  IHC staining of human colon cancer using 21206-1-AP

  Immunohistochemical analysis of paraffin-embedded human colon cancer using 21206-1-AP (HSPA4 antibody) at dilution of 1:100 (under 40x lens).

• 

  IHC staining of human breast cancer using 21206-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer using 21206-1-AP (HSPA4 antibody) at dilution of 1:100 (under 10x lens).

• 

  IHC staining of human breast cancer using 21206-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer using 21206-1-AP (HSPA4 antibody) at dilution of 1:100 (under 40x lens).

• 

  IHC staining of human pancreas cancer using 21206-1-AP

  Immunohistochemical analysis of paraffin-embedded human pancreas cancer using 21206-1-AP (HSPA4 antibody) at dilution of 1:100 (under 10x lens).

• 

  IHC staining of human pancreas cancer using 21206-1-AP

  Immunohistochemical analysis of paraffin-embedded human pancreas cancer using 21206-1-AP (HSPA4 antibody) at dilution of 1:100 (under 40x lens).

• 

  IF Staining of HepG2 using 21206-1-AP

  Immunofluorescent analysis of HepG2 cells, using HSPA4 antibody 21206-1-AP at 1:25 dilution and Rhodamine-labeled goat anti-rabbit IgG (red). Blue pseudocolor = DAPI (fluorescent DNA dye).

• 

  IF Staining of MCF-7 using 21206-1-AP

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using HSPA4 antibody (21206-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

• 

  FC experiment of C6 using 21206-1-AP

  1x10^6 C6 cells were intracellularly stained with 0.4 ug HSPA4 Polyclonal antibody (21206-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.4 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).

---

Tested Applications

• Positive WB detected in: C6 cells, HEK-293 cells, mouse testis tissue, NIH/3T3 cells, mouse brain tissue, rat brain tissue
• Positive IP detected in: mouse brain tissue
• Positive IHC detected in: human colon cancer tissue, human breast cancer tissue, human pancreas cancer 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: MCF-7 cells, HepG2 cells
• Positive FC (Intra) detected in: C6 cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:20-1:200
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 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

• WB: See 3 publications below
• ELISA: See 1 publications below

Product Information

21206-1-AP targets HSPA4 in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse, rat, monkey samples.

• Tested Reactivity: human, mouse, rat, monkey
• Cited Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: HSPA4 fusion protein Ag15581
• Full Name: heat shock 70kDa protein 4
• Calculated Molecular Weight: 840 aa, 94 kDa
• Observed Molecular Weight: 110 kDa
• GenBank Accession Number: BC110861
• Gene Symbol: HSPA4
• Gene ID (NCBI): 3308
• RRID: AB_10733641
• 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

HSPA4 (also known as Apg-2, HSP70RY) is a 110 kDa cytosolic protein, a member of the heat-shock protein 110 (HSP110) subfamily of HSP70 proteins which are highly conserved chaperons implicated in protein folding, protein refolding, protein transport, and protein targeting. HSPA4 is ubiquitously expressed and its expression is not heat inducible. Overexpression of HSPA4 has been reported in some leukemia and solid tumors. This antibody well recognized the endogenous HSPA4 protein in mouse brain/ testis tissues. (PMID: 21487003)

Publications

Species	Application	Title
mouse	WB	Int J Biochem Cell Biol; Folate deprivation induces cell cycle arrest at G0/G1 phase and apoptosis in hippocampal neuron cells through down-regulation of IGF-1 signaling pathway.; Authors - Yang Yang
human	ELISA	Eur J Clin Invest; Salivary monitoring related to major surgery.; Authors - Ullmann Yehuda Y
mouse	WB	Int J Med Sci; Exercise combined with trimetazidine improves anti-fatal stress capacity through enhancing autophagy and heat shock protein 70 of myocardium in mice.; Authors - Lingjun Jiang
human	WB	Mol Cell Proteomics; A new cellular interactome of SARS-CoV-2 nucleocapsid protein and its biological implications; Authors - Yuan-Qin Min