The first step in two-dimensional gel electrophoresis generates a series of protein bands by isoelectric focusing. In a second step a strip of this gel is turned 90 degrees, placed on another gel containing SDS, and electric current is again applied. In this second step :
(a) Proteins with similar isoelectric points become further separated according to their molecular weights.
(b) The individual bands become stained so that the isoelectric focus pattern can be visualized.
(c) The individual bands become visualized by interacting with protein-specific antibodies in the second gel.
(d) The individual bands undergo a second, more intense isoelectric focusing.
Answer:
(a) Proteins with similar isoelectric points become further separated according to their molecular weights.
Explanation:
Two-dimensional gel electrophoresis (2D-GE) is a powerful technique used to separate complex protein mixtures by employing two independent properties: isoelectric point (pI) and molecular weight (MW). The process occurs in two sequential steps:
Step 1: Isoelectric Focusing (IEF)
- Proteins are separated in a pH gradient gel based on their isoelectric point (pI), the pH at which a protein carries no net charge.
- This separation is achieved using ampholytes, which establish a stable pH gradient when an electric field is applied.
- Each protein migrates to the point in the gradient where its net charge becomes zero and remains stationary, forming discrete protein bands.
Step 2: SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis)
- The IEF gel strip is rotated 90 degrees and placed on a second gel containing SDS, a detergent that denatures proteins and imparts a uniform negative charge.
- When an electric field is applied, proteins migrate through the SDS-PAGE gel exclusively based on their molecular weight, with smaller proteins migrating faster than larger ones.
- This step further separates proteins that had the same pI but different molecular weights, leading to a more detailed resolution of complex protein mixtures.
Why Other Options Are Incorrect:
- (b) Incorrect: Staining (e.g., Coomassie Blue or silver stain) is performed after electrophoresis to visualize protein bands but is not part of the second electrophoretic step itself.
- (c) Incorrect: Protein-specific antibodies are used in Western blotting, which is a separate technique for protein identification, not in 2D-GE.
- (d) Incorrect: The second step is SDS-PAGE, not a second round of isoelectric focusing.
Conclusion:
By using isoelectric focusing followed by SDS-PAGE, 2D-GE enables highly effective protein separation. This makes it a valuable tool in proteomics, biomarker discovery, and comparative protein analysis.
87 Comments
Nisha
February 28, 2025Best website good explanation
Lokesh kumawat
February 28, 2025Done well explained
Rohit Meena
February 28, 2025Yes 👍
Abhilasha
March 1, 2025Done ✅
Shreeji Charan
March 1, 2025Nice explanation sir
Beena Meena
March 2, 2025✅👍
Shalu Choudhary
March 2, 2025Good explanation
Prami Masih
March 4, 2025Okay sir
Suman bhakar
March 3, 2025👍👍
Arushi
March 3, 2025👍👍
VIKRAM GAHLOT
March 5, 2025Ok sir
Ashok Meena
March 24, 2025Done
Monu Khan
March 29, 2025👍👍
Deepika sheoran
August 13, 2025Option A is correct answer
Protein with similar isoelectric because separated according to their molecular weight.
Neelam Sharma
August 14, 2025Proteins with similar isoelectric points become further separated according to their molecular weights
Seema
August 18, 2025A
Santosh Saini
August 14, 20252D gel=1D is hori.-IEF – charge and 2D is very. – SDS -PAGE- mass, so IEF strip rotated at 90 degree and seperate ac. to mole. Weight
Anurag Giri
August 14, 2025Proteins with similar isoelectric points become further separated according to their molecular weights.
Aman Choudhary
August 14, 2025Option A is correct
After IEF the net charge is neutral then in SDS- PAGE the separation on the basis of molecular weight.
Anjani sharma
August 14, 2025Answer 1 as in 2D gel electrophoresis firstly proteins are separated on the basis of charge and then that strip is kept in SDS medium and they are then separated on the basis of molecular weight
Dharmpal Swami
August 14, 20252d page –
1.charge
2. Mass
Karishma Don
August 14, 2025IN 2D GEL, 1ST IEF and 2nd SDS-PAGE, SO on ampholyte the proteins horizontally migrates according to the pI and further in 2nd the SDS give negative charge and the proteins of similar pI further migrates vertically according to their mol. weight
Savita Garwa
August 14, 20252D gel electrophoresis
1st charge in vertical
2nd Mass in horizontal
Mitali
August 14, 2025(a) Proteins with similar isoelectric points become further separated according to their molecular weights.
Khushi Agarwal
August 14, 2025Option a is correct
Proteins with similar isoelectric points become further separated according to their mw
Khushi Singh
August 14, 2025Done
Soniya Shekhawat
August 14, 2025Option a is correct 💯
Akshita Saini
August 14, 2025Proteins with similar isoelectric points become further separated according to their molecular weights
Urmi sisodiya
August 14, 2025Option a is correct bcoz in SDS gel the protein seperate on the basis of molecular weight
Meenakshi Choudhary
August 14, 20252d gel-
Isoelectric focusing -charge
Sds page -mass
Option a is correct
Deepika sheoran
August 14, 2025Option A is correct answer
Because protein with similar isoelectric points are then separated by molecular weight
Avni
August 14, 2025The correct option is (a) Proteins with similar isoelectric points become further separated according to their molecular weights.
Nilofar Khan
August 14, 2025Option a is correct
Roopal Sharma
August 14, 2025In 2D gel electrophoresis proteins are separated on the basis of charge and mass
Deepika Sheoran
August 14, 2025Proteins with similar isoelectric points are then separated by molecular weight using SDS PAGE.
Khushi Mehra
August 14, 2025Proteins which have similar isoelectric point are separated on the basis of their molecular weight by gel electrophoresis
Priya dhakad
August 14, 2025Proteins with similar isoelectric points become further separated according to their molecular weights.
Monika jangid
August 14, 2025In 2d protein are separate on the basis of mass so option A is correct
Mansukh Kapoor
August 14, 2025The correct answer is option 1st because the proteins with same isoelectric point separates in a series according to there molecular weight
Dipti Sharma
August 14, 2025(a) Proteins with similar isoelectric points become further separated according to their molecular weights .
Surbhi Rajawat
August 14, 2025PAGE separates the proteins on the basis of molecular weight which is the 2nd step of 2D gel and all those proteins have same Pi therefore answer is 1st
Parul Yadav
August 14, 2025During IEF, proteins are separated according to their charge and in SDS-PAGE they are further separated according to their Mol.wt.
Shivani
August 14, 2025Option (a)
Because in 2D sds gel proteins are separated on the basis of mass/ molecular weight
Ankita Pareek
August 15, 2025In 2D GE proteins first separate on the basis of charge then on the basis of molecular weight
HIMANI FAUJDAR
August 15, 2025Option (a) is correct,In 2D gel protein with same dielectric point separate on the basis mass.
Kajal
August 15, 2025All protein have the same PI then they will separate acc. To their diff molecular weight
Kavita choudhary
August 15, 2025Answer a
Devika
August 15, 2025Done sir protein separation on basis of molecular weight in vertical gel medium
shruti sharma
August 16, 2025ans a
Payal Gaur
August 17, 2025Sepration of protein in SDS gel strip is vertical and based on mass.
Aafreen
August 17, 2025Proteins with similar isoelectric points become further separated according to their molecular weight
Snehakumawat
August 17, 2025Jin proteins ka pi same hota h vo agle step me molecular weight ke according separate hote Hain
Khushi Vaishnav
August 17, 2025Proteins with similar isoelectric points become further separated according to their molecular weights
Neeraj Sharma
August 17, 2025In 2D gel electrophoresis there are two steps in first the proteins are separated on the basis of charge and in 2 step the proteins will further separated on the basis of molecular weight so option 1 os correct
Simran Saini
August 17, 2025Protein with similar isoelectric points become further separated according to their molecular weights.
Komal Sharma
August 19, 2025Two-dimensional gel electrophoresis (2D-GE) is a powerful technique used to separate complex protein mixtures by employing two independent properties: isoelectric point (pI) and molecular weight (MW). The process occurs in two sequential steps so option 1 is correct
Komal Sharma
August 19, 2025By using isoelectric focusing followed by SDS-PAGE, 2D-GE enables highly effective protein separation. This makes it a valuable tool in proteomics, biomarker discovery, and comparative protein analysis so according to this option 1 is correct
Varsha Tatla
August 23, 2025In 2d gel electrophoresis here two steps involves 1st the protein separated on the basis of charge n 2nd the protein separated on the basis of size
Bharti Yadav
August 24, 2025(a) Proteins with similar isoelectric points become further separated according to their molecular weights.
Priyanka Choudhary
August 24, 2025Correct answer is option a, because of this
Two-dimensional gel electrophoresis is a powerful technique used to separate proteins based on two properties charge and size
Urmi
August 24, 2025In 2d gel electrophoresis,If a protein have same isoelectric point then it further seperated by on the basis of molecular weight
MOHIT AKHAND
August 24, 2025Proteins with similar isoelectric points become further separated according to their molecular weights with the help of 2-D gel electrophoresis
Anil Choudhary
August 24, 2025Proteins with similar isoelectric points become further separated according to their molecular weights
Anju
August 24, 2025Answer:a
In 2D gel electrophoresis if protein has same isoelectric point then they are separated on the bases of molecular weight
Bhavana kankhedia
August 24, 2025Proteins with similar isoelectric points become further separated according to their molecular weights
Alec
August 24, 2025correct ans. is (a) because proteins with similar isoelectric points are separated according to their molecular weights.
Anjali
August 24, 2025Proteins with similar isoelectric points become further separated according to their molecular weights. So a is correct.
Sonam Saini
August 24, 2025Isoelectric foxing IEf
Sakshi
August 24, 2025Protein with similar isoelectric points become further separated according to their molecular weight
Shivani Panwar
August 24, 2025Ans is A
Devika
August 24, 2025Proteins with similar pi separate according to their molecular weight
Falguni Prakash
August 24, 2025A
Kashish yadav
August 25, 2025👍👍
sakshi khatri
August 25, 2025done
Pratibha Jain
August 30, 2025Correct answer is option (a)
Proteins with similar isoelectric points become further separated according to their molecular weights.
Shriyanshi verma
August 30, 2025Option A is correct answer
Protein with similar isoelectric because separated according to their molecular weight.
Sheikh Aleen
August 30, 2025Proteins with similar isoelectric points become further separated according to their molecular weights.
Patel Radha
August 30, 2025Well explanation
Swati Choudhary
August 31, 2025A is correct –
Proteins with similar isoelectric points become further separated according to their molecular weights.
Shobha Kanwar
August 31, 2025In 2D GE proteins first separate on the basis of charge then on the basis of molecular weight
Aartii sharma
September 1, 2025(a) protein with same isoelectric point become further separated according to their molecular weight
Ajay Sharma
September 2, 2025Protein with same isoelectric point gets seperate according to their mw
Anisha jakhar
September 3, 2025Proteins with similar isoelectric points become further separated according to their molecular weights.
Muskan Yadav
September 3, 2025Proteins with similar isoelectric points become further separated according to their molecular weights.
Somya Yadav
September 5, 2025Protein with similar iso electric point will separate on the basis of mol. Wt.
Aakansha sharma Sharma
October 1, 2025(a) Proteins with similar isoelectric points become further separated according to their molecular weights
Harish Kumar
November 18, 2025A Option is correct