RPB5 Polyclonal antibody

RPB5 Polyclonal Antibody for WB, IP, IHC, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IP, IHC, ELISA

Conjugate

Unconjugated

Publications

5

Cat no : 15217-1-AP

Synonyms

hRPB25, hsRPB5, POLR2E, RPABC1, RPB5, RPB5 homolog, XAP4

---

Validation Data Gallery

• 

  WB analysis using 15217-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 15217-1-AP (RPB5 antibody) at dilution of 1:3000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of rat liver using 15217-1-AP

  rat liver tissue were subjected to SDS PAGE followed by western blot with 15217-1-AP (RPB5 antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of rat spleen using 15217-1-AP

  rat spleen tissue were subjected to SDS PAGE followed by western blot with 15217-1-AP (RPB5 antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse spleen using 15217-1-AP

  mouse spleen tissue were subjected to SDS PAGE followed by western blot with 15217-1-AP (RPB5 antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of mouse liver using 15217-1-AP

  mouse liver tissue were subjected to SDS PAGE followed by western blot with 15217-1-AP (RPB5 antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HeLa using 15217-1-AP

  HeLa cells were subjected to SDS PAGE followed by western blot with 15217-1-AP (POLR2E antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HeLa using 15217-1-AP

  HeLa cells were subjected to SDS PAGE followed by western blot with 15217-1-AP (POLR2E antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of HepG2 using 15217-1-AP

  HepG2 cells were subjected to SDS PAGE followed by western blot with 15217-1-AP (POLR2E antibody) at dilution of 1:800 incubated at room temperature for 1.5 hours.

• 

  WB analysis of MCF-7 using 15217-1-AP

  MCF7 cells were subjected to SDS PAGE followed by western blot with 15217-1-AP (POLR2E antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of A549 using 15217-1-AP

  A549 cells were subjected to SDS PAGE followed by western blot with 15217-1-AP (POLR2E antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

• 

  IP experiment of HeLa using 15217-1-AP

  IP result of anti-RPB5 (IP:15217-1-AP, 4ug; Detection:15217-1-AP 1:500) with HeLa cells lysate 880ug.

• 

  IHC staining of human breast cancer using 15217-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 15217-1-AP (RPB5 antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 15217-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 15217-1-AP (RPB5 antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

---

Tested Applications

• Positive WB detected in: A549 cells, HeLa cells, MCF-7 cells, HepG2 cells, rat liver tissue, rat spleen tissue, mouse spleen tissue, mouse liver tissue, Jurkat cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human breast cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:6000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500

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 4 publications below

Product Information

15217-1-AP targets RPB5 in WB, IP, IHC, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: RPB5 fusion protein Ag7386
• Full Name: polymerase (RNA) II (DNA directed) polypeptide E, 25kDa
• Calculated Molecular Weight: 25 kDa
• Observed Molecular Weight: 23-25 kDa
• GenBank Accession Number: BC004441
• Gene Symbol: POLR2E
• Gene ID (NCBI): 5434
• RRID: AB_2299932
• 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

DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. Common component of RNA polymerases I, II and III which synthesize ribosomal RNA precursors, mRNA precursors and many functional non-coding RNAs, and small RNAs, such as 5S rRNA and tRNAs, respectively. polymerase (RNA) II is the central component of the basal RNA polymerase II transcription machinery. Pols are composed of mobile elements that move relative to each other. In Pol II, POLR2E (Polymerase (RNA) II (DNA directed) polypeptide E) /RPB5 is part of the lower jaw surrounding the central large cleft and thought to grab the incoming DNA template. Seems to be the major component in this process. The observed molecular weight of POLR2E is about 25 kDa.

Publications

Species	Application	Title
mouse	WB	Mol Cell; RNA Pol II preferentially regulates ribosomal protein expression by trapping disassociated subunits; Authors - Yuanjun Li
human	WB	Mol Cell; SPT5 stabilization of promoter-proximal RNA polymerase II.; Authors - Yuki Aoi
mouse	WB	Mol Cell; Targeted protein degradation reveals RNA Pol II heterogeneity and functional diversity; Authors - Yuanjun Li
human	WB	Oncogene; RMP promotes venous metastases of hepatocellular carcinoma through promoting IL-6 transcription.; Authors - J Zhang
mouse		Genome Biol; Cross-regulome profiling of RNA polymerases highlights the regulatory role of polymerase III on mRNA transcription by maintaining local chromatin architecture; Authors - Yongpeng Jiang