RB1 Antibody (607121) [Unconjugated]

Western blot shows lysates of Jurkat human acute T cell leukemia cell line, Daudi human Burkitt's lymphoma cell line, and Raji human Burkitt's lymphoma cell line. PVDF Membrane was probed with 0.1 µg/mL of Human RB1 Monoclonal Antibody (Catalog # MAB6495) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for RB1 at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

RB1 was detected in immersion fixed MCF-7 human breast cancer cell line using Human RB1 Monoclonal Antibody (Catalog # MAB6495) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for RB1 at approximately 120 kDa (as indicated) using 1 µg/mL of Mouse Anti-Human RB1 Monoclonal Antibody (Catalog # MAB6495). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Simultaneous knockdown of multiple genes in primary human islets following electroporation. (A) Graph shows RT-qPCR results of islets electroporated with non-targeting control siRNA or siRNA targeting RB and p130 (L-003299). (B) Graph shows RT-qPCR results of islets electroporated with non-targeted control siRNA or siRNAs targeting the indicated gene products. The housekeeping gene GAPDH was used as the control. Representative experiment shown from 3 separate experiments. Error bars indicate standard deviation. (C) Western blot for RB protein in human islet lysates after electroporation of control or targeting siRNA. GAPDH was used as a loading control. RB appears as a characteristic doublet representing the hypo and hyperphosphorylated protein. Representative experiment shown from 3 different experiments. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/25305068), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Simple Western, ICC/IF
• Clone: 607121
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

RB1 Antibody (607121) [Unconjugated] Summary

• Immunogen: E.coli-derived recombinant human RB1
  Lys240-Asn406
  Accession # P06400
• Specificity: Detects human RB1 in Western blots.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: RB1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 ug/mL
  • Simple Western 1 ug/mL
  • Western Blot 0.1 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for RB1 Antibody (607121) [Unconjugated]

• OSRC
• osteosarcoma
• p105-Rb
• pp110
• pRb
• RB
• RB1
• retinoblastoma 1
• retinoblastoma suspectibility protein
• retinoblastoma-associated protein

Background

Retinoblastoma 1 protein (RB-1; also retinoblastoma-associated protein, pp110, and p105-Rb) is a 110 kDa tumor suppressor gene and member of the retinoblastoma protein family.  Human RB-1 is 928 amino acids in length.  The protein contains a Pocket domain (aa 373-771), which is comprised of two other domains, domain A (aa 373-573) and domain B (aa 640-771), and a “spacer” (aa 580-639).  The Pocket domain binds to threonine-phosphorylated domain C (aa 771-928), which thereby prevents interaction with heterodimeric E2F/DP transcription factor complexes.  Human RB-1 is 90% aa identical to mouse RB-1.  RB-1 is expressed in the retina.  The underphosphorylated, active form of RB-1 interacts with E2F1 and represses its transcription activity, leading to cell cycle arrest.  Defects in RB-1 lead to the childhood cancer retinoblastoma.  

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for RB1 Antibody (MAB6495)(6)

Publications using MAB6495

• Manceau L, Richard Albert J, Lollini PL et al. Divergent transcriptional and transforming properties of PAX3-FOXO1 and PAX7-FOXO1 paralogs PLOS Genetics 2022-05-23 [PMID: 35604932]
• Manceau L, Richard Albert J, Lollini PL et al. Divergent transcriptional and transforming properties of PAX3-FOXO1 and PAX7-FOXO1 paralogs PLOS Genetics 2022-05-23 [PMID: 35604932]
• Liqin SHAO, Zhangxing SHENG, Yuefeng ZHU, Jianchao LI, Rufa MENG Effect of miR‑215 on the Expression of Tumor Suppressor Gene Rb1 in Retinoblastoma Cell Lines Iranian Journal of Public Health 2020-10-21 [PMID: 33083296]
• Yoshihisa Shimada, Yujin Kudo, Sachio Maehara, Jun Matsubayashi, Yoichi Otaki, Naohiro Kajiwara et al. Ubiquitin C‐terminal hydrolase‐L1 has prognostic relevance and is a therapeutic target for high‐grade neuroendocrine lung cancers Cancer Science 2020-02-01 [PMID: 31845438]
• Stanley Tamaki, Christopher Nye, Euan Slorach, David Scharp, Helen M Blau, Phyllis E Whiteley et al. Simultaneous silencing of multiple RB and p53 pathway members induces cell cycle reentry in intact human pancreatic islets BMC Biotechnology 2014-12-01 [PMID: 25305068]
• Stanley Tamaki, Christopher Nye, Euan Slorach, David Scharp, Helen M Blau, Phyllis E Whiteley et al. Simultaneous silencing of multiple RB and p53 pathway members induces cell cycle reentry in intact human pancreatic islets BMC Biotechnology 2014-12-01 [PMID: 25305068] (Western Blot, Human)

Bioinformatics

• Gene Symbol: RB1
• Uniprot:
  • P06400()