CBX5 Recombinant antibody

CBX5 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

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

Conjugate

Unconjugated

CloneNo.

240145F11

Cat no : 83258-5-RR

Synonyms

HP1A, HP1 alpha, HP1, Antigen p25, 240145F11

---

Validation Data Gallery

• 

  WB analysis using 83258-5-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 83258-5-RR (CBX5 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of human placenta using 83258-5-RR

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 83258-5-RR (CBX5 antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human placenta using 83258-5-RR

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 83258-5-RR (CBX5 antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse brain using 83258-5-RR

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 83258-5-RR (CBX5 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse stomach using 83258-5-RR

  Immunohistochemical analysis of paraffin-embedded mouse stomach tissue slide using 83258-5-RR (CBX5 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat kidney using 83258-5-RR

  Immunohistochemical analysis of paraffin-embedded rat kidney tissue slide using 83258-5-RR (CBX5 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat kidney using 83258-5-RR

  Immunohistochemical analysis of paraffin-embedded rat kidney tissue slide using 83258-5-RR (CBX5 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat stomach using 83258-5-RR

  Immunohistochemical analysis of paraffin-embedded rat stomach tissue slide using 83258-5-RR (CBX5 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat stomach using 83258-5-RR

  Immunohistochemical analysis of paraffin-embedded rat stomach tissue slide using 83258-5-RR (CBX5 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of HepG2 using 83258-5-RR

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using CBX5 antibody (83258-5-RR, Clone: 240145F11 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red).

• 

  FC experiment of SH-SY5Y using 83258-5-RR

  1x10^6 SH-SY5Y cells were intracellularly stained with 0.25 ug CBX5 Recombinant antibody (83258-5-RR, Clone:240145F11) and APC-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed and permeabilized with True-Nuclear Transcription Factor Buffer Set.

• 

  Affinity and Kinetic Characterization of 83258-5-RR

  Biolayer interferometry (BLl) kinetic assays of 83258-5-RR against Human CBX5 were performed. The affinity constant is 0.339 nM.

---

Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, MCF-7 cells, Jurkat cells, A431 cells
• Positive IHC detected in: human placenta tissue, mouse brain tissue, mouse stomach tissue, rat kidney tissue, rat stomach 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: HepG2 cells
• Positive FC (Intra) detected in: SH-SY5Y cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 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

83258-5-RR targets CBX5 in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: CBX5 fusion protein Ag34810
• Full Name: chromobox homolog 5 (HP1 alpha homolog, Drosophila)
• Calculated Molecular Weight: 191 aa, 22 kDa
• Observed Molecular Weight: 22 kDa
• GenBank Accession Number: BC006821
• Gene Symbol: CBX5
• Gene ID (NCBI): 23468
• 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

Chromobox protein homolog 5 (CBX5), also named heterochromatin protein 1 alpha (HP1a), is a highly conserved nonhistone protein involved in heterochromatin formation and gene silencing in different species including humans.HP1a is a Component of heterochromatin that recognizes and binds histone H3 tails methylated at 'Lys-9' (H3K9me), leading to epigenetic repression. In contrast, it is excluded from chromatin when 'Tyr-41' of histone H3 is phosphorylated (H3Y41ph). It may interact with lamin-Breceptor. HP1a is involved in the formation of functional kinetochore through interaction with MIS12complex proteins. Phosphorylation of HP1 and LBR during interphase mitosis may be responsible for some of the alterations in chromatin organization and nuclear structure which occur at various times during the cell cycle.The HP1a was expressed in nucleus and associates specifically with chromatin during metaphase and anaphase.Recent studies have shown that HP1a is present at many euchromatic sites and positively regulates euchromatic gene expression through RNA transcript association and interaction with hnRNPs in Drosophila (19798443).