HSPB3 Monoclonal antibody

HSPB3 Monoclonal Antibody for WB, IF, FC, IHC, ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

Human, rabbit, mouse, rat

Applications

WB, IF, FC, IHC, ELISA

Conjugate

Unconjugated

CloneNo.

2A10B5

Cat no : 67890-1-Ig

Synonyms

Heat shock 17 kDa protein, heat shock 27kDa protein 3, Heat shock protein beta 3, HSP 17, HSPB3, HSPL27

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Validation Data Gallery

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  WB analysis using 67890-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 67890-1-Ig (HSPB3 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated GAPDH Monoclonal antibody (HRP-60004) as loading control.

• 

  WB analysis of human heart using 67890-1-Ig

  human heart tissue were subjected to SDS PAGE followed by western blot with 67890-1-Ig (HSPB3 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse heart using 67890-1-Ig

  mouse heart tissue were subjected to SDS PAGE followed by western blot with 67890-1-Ig (HSPB3 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of rabbit heart using 67890-1-Ig

  rabbit heart tissue were subjected to SDS PAGE followed by western blot with 67890-1-Ig (HSPB3 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of rat kidney using 67890-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat kidney tissue slide using 67890-1-Ig (HSPB3 antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse kidney using 67890-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 67890-1-Ig (HSPB3 antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse kidney using 67890-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 67890-1-Ig (HSPB3 antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse kidney using 67890-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 67890-1-Ig (HSPB3 antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat kidney using 67890-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat kidney tissue slide using 67890-1-Ig (HSPB3 antibody) at dilution of 1:500 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat kidney using 67890-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat kidney tissue slide using 67890-1-Ig (HSPB3 antibody) at dilution of 1:500 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HeLa using 67890-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using HSPB3 antibody (67890-1-Ig, Clone: 2A10B5 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• 

  IF Staining of HeLa using 67890-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using HSPB3 antibody (67890-1-Ig, Clone: 2A10B5 ) at dilution of 1:800 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

• 

  FC experiment of HeLa using 67890-1-Ig

  1X10^6 HeLa cells were intracellularly stained with 0.5 ug Anti-Human HSPB3 (67890-1-Ig, Clone:2A10B5) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.5 ug Mouse IgG2b Isotype Control (66360-3-Ig, Clone: K11B8C4B5) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: human heart tissue, mouse heart tissue, rabbit heart tissue, rat heart tissue
• Positive IHC detected in: rat kidney tissue, mouse kidney 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
• Positive FC detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:250-1:1000
• Immunofluorescence (IF): IF : 1:400-1:1600
• Flow Cytometry (FC): FC : 0.50 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.

Product Information

67890-1-Ig targets HSPB3 in WB, IF, FC, IHC, ELISA applications and shows reactivity with Human, rabbit, mouse, rat samples.

• Tested Reactivity: Human, rabbit, mouse, rat
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: HSPB3 fusion protein Ag26940
• Full Name: heat shock 27kDa protein 3
• Calculated Molecular Weight: 150 aa, 17 kDa
• Observed Molecular Weight: 20 kDa
• GenBank Accession Number: BC126272
• Gene Symbol: HSPB3
• Gene ID (NCBI): 8988
• RRID: AB_2918646
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A 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

Heat shock protein family B (small) member 3 (HSPB3) is the third member of the sHSP family in human and is mainly expressed in skeletal and smooth muscles, and can associate with HSPB2, which is another sHSP prominently expressed in cardiac and skeletal muscles. HSPB3 is involved in several cellular functions such as cell signaling, differentiation, and apoptosis, and mutations of HSPB3 are associated with neuromuscular diseases, such as axonal Charcot‐Marie‐Tooth disease (PMID: 31709619) (PMID: 32093037). The molecular weight of HSPB3 is about 17 kDa.