JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB273708

Human RXRA knockout HCT116 cell line

Be the first to review this product! Submit a review

|

(0 Publication)

RXRA KO cell line available to order. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

View Alternative Names

FLJ00280, FLJ00318, FLJ16020, FLJ16733, MGC102720, NR2B1, Nuclear receptor subfamily 2 group B member 1, OTTHUMP00000022510, RXR alpha1, RXRA_HUMAN, Retinoic acid receptor RXR-alpha, Retinoid X nuclear receptor alpha, Retinoid X receptor alpha

4 Images
Immunocytochemistry/ Immunofluorescence - Human RXRA knockout HCT116 cell line (AB273708)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Human RXRA knockout HCT116 cell line (AB273708)

ab125001 staining Retinoid X Receptor alpha in wild-type HCT116 cells (top panel) and RXRA knockout HCT116 cells (bottom panel) (ab273708). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab125001 at 0.2μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI. Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

Western blot - Human RXRA knockout HCT116 cell line (AB273708)
  • WB

Unknown

Western blot - Human RXRA knockout HCT116 cell line (AB273708)

Lanes 1 - 4 : Merged signal (red and green). Green - ab125001 observed at 53 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab125001 was shown to react with Anti-Retinoid X Receptor alpha in wild-type HCT 116 cells in western blot with loss of signal observed in RXRA knockout cell line ab273708 (RXRA knockout cell lysate ab275245). Wild-type and RXRA knockout HCT 116 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab125001 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (<a href='/en-us/products/primary-antibodies/retinoid-x-receptor-alpha-rxra-antibody-epr7106-ab125001'>ab125001</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT116 cell lysate at 20 µg

Lane 2:

RXRA knockout HCT116 cell lysate at 20 µg

Lane 2:

Western blot - Human RXRA knockout HCT116 cell line (ab273708)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Predicted band size: 51 kDa

Observed band size: 53 kDa

false

Sanger Sequencing - Human RXRA knockout HCT116 cell line (AB273708)
  • Sanger seq

Unknown

Sanger Sequencing - Human RXRA knockout HCT116 cell line (AB273708)

Allele-1 : 1 bp insertion in exon 1.

Western blot - Human RXRA knockout HCT116 cell line (AB273708)
  • WB

Lab

Western blot - Human RXRA knockout HCT116 cell line (AB273708)

All lanes:

Western blot - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (<a href='/en-us/products/primary-antibodies/retinoid-x-receptor-alpha-rxra-antibody-epr7106-ab125001'>ab125001</a>) at 1/500 dilution

Lane 1:

Wild-type HCT116 lysate at 20 µg

Lane 2:

Western blot - Human RXRA knockout HCT116 cell line (ab273708) at 20 µg

Observed band size: 51 kDa

false

Key facts

Cell type

HCT116

Species or organism

Human

Tissue

Colon

Form

Liquid

form

Knockout validation

Immunocytochemistry,Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1

Antibiotic resistance

Puromycin 1µg/mL

Disease

Carcinoma

style
left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "ICC/IF": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
style
left-column

Product details

We will provide viable cells that proliferate on revival.

This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

style
left-column

What's included?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab273708-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab273708 Human RXRA knockout HCT116 cell line", "number":"AB273708-CMP01" }, { "size":"1 x 1000000 Cells/vial", "name":"ab273730 Human wild-type HCT116 cell line", "number":"AB273708-CMP02" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab273708-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab273708 Human RXRA knockout HCT116 cell line", "number":"AB273708-CMP01", "productcode":"" } ] } } }
style
left-column

Properties and storage information

Gene name
RXRA
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Immunocytochemistry, Sanger Sequencing, Western blot
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
style
left-column

Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be passaged when they have achieved 80-90% confluence.
Culture medium

McCoY5a + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

style
left-column

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Retinoid X Receptor alpha (RXRA) also referred to as NR2B1 is a nuclear receptor with a mass of approximately 55 kDa. RXRA functions as a transcription factor and plays a significant role in regulating gene expression by forming heterodimers with other nuclear receptors. RXRA is expressed in many tissues with notable levels in organs like the liver lungs and kidneys. Its ability to bind DNA sequences known as retinoic acid response elements allows RXRA to regulate diverse physiological processes.
Biological function summary

RXRA serves an important role in metabolic regulation especially in lipid metabolism and glucose homeostasis. It forms heterodimers with partners such as peroxisome proliferator-activated receptors (PPARs) enhancing its capacity to modulate various metabolic pathways. These heterodimers facilitate the transcription of genes that respond to nutritional changes. RXRA also influences immune responses and cell differentiation making it important for maintaining cellular health.

Pathways

RXRA operates within the retinoid signaling and lipid metabolism pathways. Through its interaction with PPARs particularly PPAR-γ and PPAR-α RXRA becomes an integral part of the signaling that controls fatty acid storage and glucose metabolism. These pathways influence cellular function and energy balance impacting the body's overall metabolic status.

RXRA has associations with metabolic diseases such as diabetes and cardiovascular disorders. Dysregulation of RXRA expression or function can disrupt lipid and glucose metabolism leading to insulin resistance and metabolic syndrome. The RXRA-PPAR-γ interaction is essential in these conditions with the pair influencing adipogenesis and inflammatory responses. Understanding and targeting RXRA interactions can provide therapeutic prospects for these metabolic disorders.
style
left-column
style
left-column

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

style
left-column

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

style
left-column

Product protocols

style
left-column

Target data

See full target information RXRA
style
left-column
style
left-column
style
right-column

Complementary Products

Primary Antibodies

AB275745

Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated|20ul selling size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Retinoic Acid Receptor alpha antibody [EPR23871-271] (AB275745)

  • 4
  • 1
Cell Lines & Lysates

AB275245

Human RXRA knockout HCT116 cell lysate

Western blot - Human RXRA knockout HCT116 cell lysate (AB275245)

  • 0
  • 0
Primary Antibodies

AB125001

Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106]

RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-Retinoid X Receptor alpha/RXRA antibody [EPR7106] (AB125001)

  • 5
  • 5
Cell Lines & Lysates

AB266955

Human ATF3 knockout A549 cell line

Advanced Validation

Western blot - Human ATF3 knockout A549 cell line (AB266955)

  • 0
  • 0

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com