Log In Sign up
The largest database of trusted experimental protocols

Ab64575

Manufactured by Abcam
Visit Supplier
Sourced in United Kingdom

Ab64575 is a primary antibody produced in rabbit. It is designed for use in western blotting applications.

Automatically generated - may contain errors

Lab products found in correlation

Pierce ecl, by Thermo Fisher Scientific (3 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Ab181602, by Abcam (1 mentions) Ab75898, by Abcam (1 mentions) Ab17721, by Abcam (1 mentions) Ab19808, by Abcam (1 mentions) Ab2413, by Abcam (1 mentions) Sc 8422, by Santa Cruz Biotechnology (1 mentions) Sc 28348, by Santa Cruz Biotechnology (1 mentions) Sc 5565, by Santa Cruz Biotechnology (1 mentions) Sc 8035, by Santa Cruz Biotechnology (1 mentions) Hrp conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions) Ab214486, by Abcam (1 mentions) V9131, by Merck Group (1 mentions) Immobilon pvdf membrane, by Merck Group (1 mentions) Anti rabbit and anti mouse igg, by GE Healthcare (1 mentions) Ma1 80216, by Thermo Fisher Scientific (1 mentions) Sc 544, by Santa Cruz Biotechnology (1 mentions) Xcell 2 blot module, by Thermo Fisher Scientific (1 mentions) Nupage bis tris gel, by Thermo Fisher Scientific (1 mentions)

7 protocols using ab64575

1

Western Blot for Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
The extracted protein was separated by 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and then electro-transferred to the polyvinylidene fluoride membrane. After blocking with 5% interaction of bovine at room temperature for 2 h, the membrane was incubated with diluted primary antibody [JMJD6 (1:1000, ab64575, Abcam), LSD1 (1:1200, ab17721, Abcam), ERK2 (MAPK1) (1:1000, 9926, CST) and BRD4 (1:1500, ab75898, Abcam)] at 4 °C overnight. HRP-conjugated corresponding secondary antibody (goat anti-rabbit proteintech SA00001-1; goat anti-mouse proteintech SA00001-2) was added to incubate with membrane at room temperature for 1 h. Further, the membrane was exposed to enhanced chemiluminescence (EMD Millipore Corporation, Billerica, MA) and immunoblots were visualized and captured by Bio-Rad imaging system. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:1000, ab181602, Abcam) was used as an internal reference, and the protein band image was analyzed by Image Lab software.
Pei R., Zhao L., Ding Y., Su Z., Li D., Zhu S., Xu L., Zhao W, & Zhou W. (2023). JMJD6–BRD4 complex stimulates lncRNA HOTAIR transcription by binding to the promoter region of HOTAIR and induces radioresistance in liver cancer stem cells. Journal of Translational Medicine, 21, 752.
+ Open protocol
+ Expand
2

BRD4 and JMJD6 Protein Interactions

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein was extracted from CHP134 cells and co-immunoprecipitation was carried out using a rabbit anti-BRD4 antibody (Bethyl Laboratories, Montgomery, TX), rabbit anti-JMJD6 antibody (Abcam ab135066) or control IgG as a negative control, followed by immunoblot analysis with anti-BRD4 and anti-JMJD6 antibodies. Protein co-immunoprecipitation was also carried out with an anti-N-Myc antibody (Santa Cruz Biotechnology B8.4.b) or control IgG as a negative control, followed by immunoblot analysis with the anti-JMJD6 (Abcam Ab64575) and an anti-N-Myc antibodies.
Wong M., Sun Y., Xi Z., Milazzo G., Poulos R.C., Bartenhagen C., Bell J.L., Mayoh C., Ho N., Tee A.E., Chen X., Li Y., Ciaccio R., Liu P.Y., Jiang C.C., Lan Q., Jayatilleke N., Cheung B.B., Haber M., Norris M.D., Zhang X.D., Marshall G.M., Wang J.Y., Hüttelmaier S., Fischer M., Wong J.W., Xu H., Perini G., Dong Q., George R.E, & Liu T. (2019). JMJD6 is a tumorigenic factor and therapeutic target in neuroblastoma. Nature Communications, 10, 3319.
+ Open protocol
+ Expand
3

Immunohistochemical Analysis of JMJD6 in Gliomas

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Briefly, deidentified tissue microarrays (TMAs) were constructed from gliomas after obtaining University of Kentucky Institutional Review Board Approval. Three 2-mm diameter cores per tumour were obtained, with each core embedded in a separate TMA block. A total of 104 cases comprised the TMAs, including 9 nonneoplastic controls (cortical dysplasias), 9 grade II astrocytomas, 11 grade III astrocytomas, 12 anaplastic oligodendrogliomas, 16 grade II oligodendrogliomas, and 47 grade IV glioblastomas (GBMs). Immunohistochemistry was performed for each core as described previously37 (link), but using an antibody towards JMJD6 (Abcam, ab64575). Briefly, each core was semiquantified on a relative scale from 0 to 3, with 0 = negative and 3 = strongest. Results from all 3 cores were averaged together to produce a final score for a tumour. Results were plotted based on WHO grade and differences were calculated via Mann-Whitney-Wilcoxon Test.
Miller T.E., Liau B.B., Wallace L.C., Morton A.R., Xie Q., Dixit D., Factor D.C., Kim L.J., Morrow J.J., Wu Q., Mack S.C., Hubert C.G., Gillespie S.M., Flavahan W.A., Hoffmann T., Thummalapalli R., Hemann M.T., Paddison P.J., Horbinski C.M., Zuber J., Scacheri P.C., Bernstein B.E., Tesar P.J, & Rich J.N. (2017). Transcription elongation factors represent in vivo cancer dependencies in glioblastoma. Nature, 547(7663), 355-359.
+ Open protocol
+ Expand
4

Immunohistochemical Analysis of JMJD6 in Gliomas

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Briefly, deidentified tissue microarrays (TMAs) were constructed from gliomas after obtaining University of Kentucky Institutional Review Board Approval. Three 2-mm diameter cores per tumour were obtained, with each core embedded in a separate TMA block. A total of 104 cases comprised the TMAs, including 9 nonneoplastic controls (cortical dysplasias), 9 grade II astrocytomas, 11 grade III astrocytomas, 12 anaplastic oligodendrogliomas, 16 grade II oligodendrogliomas, and 47 grade IV glioblastomas (GBMs). Immunohistochemistry was performed for each core as described previously37 (link), but using an antibody towards JMJD6 (Abcam, ab64575). Briefly, each core was semiquantified on a relative scale from 0 to 3, with 0 = negative and 3 = strongest. Results from all 3 cores were averaged together to produce a final score for a tumour. Results were plotted based on WHO grade and differences were calculated via Mann-Whitney-Wilcoxon Test.
Miller T.E., Liau B.B., Wallace L.C., Morton A.R., Xie Q., Dixit D., Factor D.C., Kim L.J., Morrow J.J., Wu Q., Mack S.C., Hubert C.G., Gillespie S.M., Flavahan W.A., Hoffmann T., Thummalapalli R., Hemann M.T., Paddison P.J., Horbinski C.M., Zuber J., Scacheri P.C., Bernstein B.E., Tesar P.J, & Rich J.N. (2017). Transcription elongation factors represent in vivo cancer dependencies in glioblastoma. Nature, 547(7663), 355-359.
+ Open protocol
+ Expand
5

Antibody Panel for IF and WB

Check if the same lab product or an alternative is used in the 5 most similar protocols
Primary antibodies employed in IF and WB include mouse monoclonal anti-β-actin (ACTB) (sc-477778, 1:1,000, Santa Cruz), mouse monoclonal anti-integrin α5β1 (MAB1999, 1:200, Millipore Sigma), mouse monoclonal anti-α-tubulin (sc-8035, 1:1,000, Santa Cruz), mouse monoclonal anti-Calreticulin (CALR) (ab22683, 1:300, Santa Cruz Biotechnology), mouse monoclonal anti-CK7 (sc-70936, 1:200, Santa Cruz), rabbit polyclonal anti-collagen IV (COL IV) (ab19808, 1:200, Abcam), mouse monoclonal anti-Fibronectin (sc-8422, 1:1,000, Santa Cruz Biotechnology), rabbit polyclonal anti-Fibronectin (ab2413, 1:2,000, Abcam), mouse monoclonal anti-HLAG (ab52454, 1:200, Abcam), mouse monoclonal anti-JMJD6 (sc-28348, 1:500, Santa Cruz), rabbit polyclonal anti-JMJD6 (ab64575, 1:1,000, Abcam), rabbit monoclonal anti-Syntaxin 6 (CB4B2 #2869. 1:50, Cell Signaling Technology), and rabbit polyclonal anti-Vimentin (sc-5565, 1:200, Santa Cruz). HRP-conjugated secondary antibodies were obtained from Santa Cruz Biotechnology and used at a concentration of 1:2,000. WBs were scanned using a CanoScanLiDE20 image scanner (Canon Canada Inc. Mississauga, ON) and analyzed for densitometry.
Alahari S., Farrell A., Ermini L., Park C., Sallais J., Roberts S., Gillmore T., Litvack M., Post M, & Caniggia I. (2021). JMJD6 Dysfunction Due to Iron Deficiency in Preeclampsia Disrupts Fibronectin Homeostasis Resulting in Diminished Trophoblast Migration. Frontiers in Cell and Developmental Biology, 9, 652607.
+ Open protocol
+ Expand
6

Western Blot Analysis of Protein Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols
For Western blot assay, cells were harvested, washed with PBS, boiled for 10 minutes in lysis buffer (125 mmol/L Tris HCl pH 6.8, 5% of SDS) and supplemented with protease inhibitors (11873580001, cOmplete, Roche). Samples were then sonicated for 20 seconds, followed by centrifugation for 15 minutes at 13,000 rpm and quantified with micro BCA Assay kit reagent (23235, Thermo Fisher Scientific) using the Spark microplate reader (TECAN). Proteins were separated by SDS-PAGE on precast 4% to 12% Bis-Tris NuPAGE gels (NP0321PK2, Thermo Fisher Scientific) and blotted to Immobilon PVDF membranes (IPVH00010, Merck Millipore) using the XCell II blot module (EI9051, Thermo Fisher Scientific). Membranes were then saturated for 1 hour in Tris-buffered saline containing 5% BSA and incubated overnight with the antihuman primary antibodies recognizing: JMJD6 (ab64575, Abcam and P1495, Sigma-Aldrich), Actin-β (A1978, Sigma-Aldrich), ERα (sc-544, G20, Santa Cruz Biotechnology and MA1–80216, Invitrogen), ANXA1 (ab214486, Abcam) and Vinculin (V9131, Sigma-Aldrich). After hybridization with the appropriate secondary antibody (anti-rabbit and anti-mouse IgG, GE HealthCare), proteins were detected by chemiluminescence (EuroClone).
Cioni B., Ratti S., Piva A., Tripodi I., Milani M., Menichetti F., Langella T., Botti L., De Cecco L., Chiodoni C., Lecis D, & Colombo M.P. (2023). JMJD6 Shapes a Pro-tumor Microenvironment via ANXA1-Dependent Macrophage Polarization in Breast Cancer. Molecular Cancer Research, 21(6), 614-627.
+ Open protocol
+ Expand
7

Quantifying JMJD6 Protein Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols
To analyze changes of JMJD6 in protein levels, total protein was first isolated from transfected A549 and H460 cells using RIPA lysis buffer (Sigma-Aldrich, Germany) Protein was quantified using BCA protein assay kit (Thermo Scientific, MA, USA). Thereafter, 10% SDS-PAGE (Beyotime, Shanghai, China) was used for isolation of protein through electrophoresis followed by protein transferring into PVDF membranes. Primary antibodies anti-JMJD6 (1:1000, ab64575, Abcam, Cambridge, UK) and anti-GAPDH (1:2000, ab8245, Abcam) were incubated at 4℃ overnight with membranes, which were first blocked by 5% non-fat milk powder. Next, PVDF membranes were rinsed with TBST (Thermo Scientific) twice and cultured with HRP-marked goat anti-rabbit IgG (1:800, ab6721, Abcam) for 2h at 25°C. Pierce™ ECL (Thermo Scientific) was used for developing and proteins were analyzed through Image J (National Institutes of Health, USA). Experiments were run in a triplicate.
Yu X., Jiang Y., Hu X, & Ge X. (2021). LINC00839/miR-519d-3p/JMJD6 axis modulated cell viability, apoptosis, migration and invasiveness of lung cancer cells. Folia histochemica et cytobiologica, 59(4).
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!