Pure antibody in western blot is showing three bands. What does these bands represent?

Hi Everyone, I have tried to run pure antibody (55kD) in 10% SDS/PAGE. I have seen three bands at 250 kD, 75 kD and 50 kD respectively. There is no reducing agent added in antibody. I'm wondering...

08 September 2017 7,860 11 View

How to measure NLRP3 inflammasome activation?

How can I measure NLRP3 inflammasoem activation? In most of the papers, I found protein and mRNA expression levels of NLRP3 and caspase-1. Since NLRP3, Asc and procaspase-1 make an assembly and it...

02 March 2017 2,472 5 View

Can anyone suggest to me where i can buy N9 microglia cell line?

Can anyone suggest to me where i can buy N9 microglia cell line?

10 November 2016 7,790 2 View

Maximum concentration of Tween-20 in TBS-T in immunoblot??

Maximum concentration of Tween-20 in TBS-T in immunoblot technique??

08 September 2016 6,281 3 View

Fuzzy and two bands instead of single crisp band in SDS/PAGE?

I'm looking for 12 kD protein expression by SDS/PAGE. The bands I was observing earlier were single crisp bands. Few days back I have started seeing fuzzy bands. I didn't make any changes in...

04 May 2016 4,729 9 View

How can I detect total s-nitrosylation in cell culture?

I'm trying to detect total S-nitrosylation in SH-SY5Y cell culture using Sodium Nitroprusside (200, 400uM). I've treated cells with SNP for 15 min and isolated proteins using lysis buffer...

31 December 2015 9,977 0 View

Can anyone suggest NAtive PAGE gel conditions?

Please correct me if you have tried any other method or reagents. 1) 12% separating gel (5ml): 30% acrylamide: 4ml, 0.375 M Tris-HCl (pH=8.8): 5.89 ml, 10% APS = 100 ul, TEMED= 10ul 2) Stacking...

08 September 2015 751 5 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

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02 March 2021 3,060 3 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View