Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
Overview
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Product name
Rabbit IgG, polyclonal - Isotype Control (ChIP Grade)
See all Rabbit isotype controls -
Tested applications
Suitable for: WB, ChIP, Flow Cytmore details -
General notes
For more information regarding the isotype control selection, please see https://www.abcam.com/primary-antibodies/your-guide-to-selecting-an-isotype-control
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches with a concentration less than 1 mg/ml will have 1% BSA -
Concentration information loading... -
Purity
Protein G purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
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Alternative names
- rabbit isotype control
Images
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Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab171870 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
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Flow Cytometry - Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)Overlay histogram showing HeLa cells stained with ab75186 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab75186, 0.05μg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150081) at 1/4000 dilution for 30 min at 22ºC.
Isotype control antibody (black line) was rabbit IgG (polyclonal) (ab171870, 0.05μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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Western blot - Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)Lanes 1-6 : Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870) at 1 µg/ml
Lanes 7-12 : Anti-beta Actin antibody (ab8227) at 1 µg/ml
Lanes 1 & 7 : Human liver tissue lysate - total protein (ab29889)
Lanes 2 & 8 : Liver (Mouse) Tissue Lysate
Lanes 3 & 9 : Liver (Rat) Tissue Lysate
Lanes 4 & 10 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lanes 5 & 11 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
Lanes 6 & 12 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Exposure time: 2 minutesPlease note that ab171870 in lanes 1-6 represents a negative control for Beta Actin, positively seen in lanes 7-12.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab171870 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
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Western blot - Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)All lanes : Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870) at 1 µg/ml
Lane 1 : Liver (Human) Tissue Lysate - adult normal tissue
Lane 2 : Brain (Mouse) Tissue Lysate
Lane 3 : Rat Kidney Tissue Lysate
Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 5 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 6 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Exposure time: 20 minutesThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab171870 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.
