#loading control

How to select a proper loading control antibody for Western Blot?

Loading control is also called internal control. In terms of mammalian cell expression, it generally refers to the protein encoded by housekeeping genes (Housekeeping Proteins). Their expression in various tissues and cells is relatively stable. It is often used as a reference while testing the expression level of a specific protein. In the Western Blotting experiment, it is proposed to do a internal control detection which can correct protein quantification and make up for the experiment error in loading samples procedure, ensuing the accuracy of the experimental results.

  There are two kinds of method about how to use loading control antibody in Western Blot.

Conjugated loading control antibody detection: Add HRP conjugated loading control antibody in secondary antibody incubation.

Normal loading control antibody detection

If molecular weight(MW) is similar between your targeted protein and selected loading control protein, it is proposed to do the detection of target protein firstly, then using strip buffer to wash the antibody in membrane, and doing the detection of loading control antibody.

If molecular weight(MW)difference is obvious between your targeted protein and selected loading control protein, using prestained protein marker after transmembrane, according to the size of protein marker, then cut off the membrane into two parts, high molecular weight and low molecular weight, dividing your target protein and loading control protein. Finally do Incubation in separate membrane for your primary antibody and loading control antibody, secondary antibody incubation and color development.

There are many kinds of loading control, so choose a right loading control for your experiment is very important. Below points can help you to better make the decision.

Sample species: For tissues and cells of mammals, it is generally recommended to choose the proteins like beta-actin, Tubulin, GAPDH, Lamin B, PCNA, Na+/K+-ATPase, etc. Plant actin and Rubisco are often used in plant samples.

Molecular weight of the target protein: Select a loading control that has a different molecular weight with the protein of interest (>5 kDa), so that you could distinguish between the bands. For high molecular weight (>100 kDa) proteins, we recommend Vinculin as a loading control.

Expression location of the target protein: whole cell, nuclear, mitochondrial, membrane, etc..

In some certain conditions, the expression of loading controls will be abnormal in some cells, this should be taken into account.

  Abbkine offers a wide range of loading control antibodies, including the hot targets and unique species, covering the molecular weights from 15 kDa to 116 kDa. Also our conjugated loading control antibodies have diverse conjugates, including enzyme and featured fluorescent dyes, with the application of WB, IHC, IF and IP experiments.

  Taking a specific example here, if you extract total proteins or cytoplasmic proteins from cells, beta-actin, GAPDH and Tubulin will be your choice, and luckily Abbkine can provide a series of antibodies helpful for your research, kindly refer to below:

CAT# Product Name MW Application Publications A01010 Anti-β-Actin Mouse Mab (1C7) 43 kDa IF, IHC, WB

  48 A01020 Anti-GAPDH Mouse Mab (2B5) 37 kDa IF, IHC, WB 47 A01030 Anti-β-Tubulin Mouse Mab (3G6) 55 kDa IF, IHC, WB 8 A01080 Anti-α-Tubulin Mouse Mab (3G5) 55 kDa WB, IHC, IF, IP A01130 Anti-Cyclophilin B Mouse Mab (7B2) 24 kDa WB, IHC, IF

  If you have any further interests to learn Abbkine loading control antibody category, kindly refer at https://www.abbkine.com/product-category/antibody/loading-control/ , send an E-mail to [email protected], our representative will surely contact you back shortly.

  About Abbkine Scientific Co., Ltd.

How to select a proper loading control antibody for Western Blot?

Loading control is also called internal control. In terms of mammalian cell expression, it generally refers to the protein encoded by housekeeping genes (Housekeeping Proteins). Their expression in various tissues and cells is relatively stable. It is often used as a reference while testing the expression level of a specific protein. In the Western Blotting experiment, it is proposed to do a internal control detection which can correct protein quantification and make up for the experiment error in loading samples procedure, ensuing the accuracy of the experimental results.

  There are two kinds of method about how to use loading control antibody in Western Blot.

Conjugated loading control antibody detection: Add HRP conjugated loading control antibody in secondary antibody incubation.

Normal loading control antibody detection

If molecular weight(MW) is similar between your targeted protein and selected loading control protein, it is proposed to do the detection of target protein firstly, then using strip buffer to wash the antibody in membrane, and doing the detection of loading control antibody.

If molecular weight(MW)difference is obvious between your targeted protein and selected loading control protein, using prestained protein marker after transmembrane, according to the size of protein marker, then cut off the membrane into two parts, high molecular weight and low molecular weight, dividing your target protein and loading control protein. Finally do Incubation in separate membrane for your primary antibody and loading control antibody, secondary antibody incubation and color development.

There are many kinds of loading control, so choose a right loading control for your experiment is very important. Below points can help you to better make the decision.

Sample species: For tissues and cells of mammals, it is generally recommended to choose the proteins like beta-actin, Tubulin, GAPDH, Lamin B, PCNA, Na+/K+-ATPase, etc. Plant actin and Rubisco are often used in plant samples.

Molecular weight of the target protein: Select a loading control that has a different molecular weight with the protein of interest (>5 kDa), so that you could distinguish between the bands. For high molecular weight (>100 kDa) proteins, we recommend Vinculin as a loading control.

Expression location of the target protein: whole cell, nuclear, mitochondrial, membrane, etc..

In some certain conditions, the expression of loading controls will be abnormal in some cells, this should be taken into account.

  Abbkine offers a wide range of loading control antibodies, including the hot targets and unique species, covering the molecular weights from 15 kDa to 116 kDa. Also our conjugated loading control antibodies have diverse conjugates, including enzyme and featured fluorescent dyes, with the application of WB, IHC, IF and IP experiments.

  Taking a specific example here, if you extract total proteins or cytoplasmic proteins from cells, beta-actin, GAPDH and Tubulin will be your choice, and luckily Abbkine can provide a series of antibodies helpful for your research, kindly refer to below:

CAT# Product Name MW Application Publications A01010 Anti-β-Actin Mouse Mab (1C7) 43 kDa IF, IHC, WB

  48 A01020 Anti-GAPDH Mouse Mab (2B5) 37 kDa IF, IHC, WB 47 A01030 Anti-β-Tubulin Mouse Mab (3G6) 55 kDa IF, IHC, WB 8 A01080 Anti-α-Tubulin Mouse Mab (3G5) 55 kDa WB, IHC, IF, IP A01130 Anti-Cyclophilin B Mouse Mab (7B2) 24 kDa WB, IHC, IF

  If you have any further interests to learn Abbkine loading control antibody category, kindly refer at https://www.abbkine.com/product-category/antibody/loading-control/ , send an E-mail to [email protected], our representative will surely contact you back shortly.

  About Abbkine Scientific Co., Ltd.

How to select a proper loading control antibody for Western Blot?

  Loading control is also called internal control. In terms of mammalian cell expression, it generally refers to the protein encoded by housekeeping genes (Housekeeping Proteins). Their expression in various tissues and cells is relatively stable. It is often used as a reference while testing the expression level of a specific protein. In the Western Blotting experiment, it is proposed to do a internal control detection which can correct protein quantification and make up for the experiment error in loading samples procedure, ensuing the accuracy of the experimental results.

  There are two kinds of method about how to use loading control antibody in Western Blot.

Conjugated loading control antibody detection: Add HRP conjugated loading control antibody in secondary antibody incubation.

Normal loading control antibody detection

If molecular weight(MW) is similar between your targeted protein and selected loading control protein, it is proposed to do the detection of target protein firstly, then using strip buffer to wash the antibody in membrane, and doing the detection of loading control antibody.

If molecular weight(MW)difference is obvious between your targeted protein and selected loading control protein, using prestained protein marker after transmembrane, according to the size of protein marker, then cut off the membrane into two parts, high molecular weight and low molecular weight, dividing your target protein and loading control protein. Finally do Incubation in separate membrane for your primary antibody and loading control antibody, secondary antibody incubation and color development.

There are many kinds of loading control, so choose a right loading control for your experiment is very important. Below points can help you to better make the decision.

Sample species: For tissues and cells of mammals, it is generally recommended to choose the proteins like beta-actin, Tubulin, GAPDH, Lamin B, PCNA, Na+/K+-ATPase, etc. Plant actin and Rubisco are often used in plant samples.

Molecular weight of the target protein: Select a loading control that has a different molecular weight with the protein of interest (>5 kDa), so that you could distinguish between the bands. For high molecular weight (>100 kDa) proteins, we recommend Vinculin as a loading control.

Expression location of the target protein: whole cell, nuclear, mitochondrial, membrane, etc..

In some certain conditions, the expression of loading controls will be abnormal in some cells, this should be taken into account.

  Abbkine offers a wide range of loading control antibodies, including the hot targets and unique species, covering the molecular weights from 15 kDa to 116 kDa. Also our conjugated loading control antibodies have diverse conjugates, including enzyme and featured fluorescent dyes, with the application of WB, IHC, IF and IP experiments.

  Taking a specific example here, if you extract total proteins or cytoplasmic proteins from cells, beta-actin, GAPDH and Tubulin will be your choice, and luckily Abbkine can provide a series of antibodies helpful for your research, kindly refer to below:

CAT# Product Name MW Application Publications A01010 Anti-β-Actin Mouse Mab (1C7) 43 kDa IF, IHC, WB

  48 A01020 Anti-GAPDH Mouse Mab (2B5) 37 kDa IF, IHC, WB 47 A01030 Anti-β-Tubulin Mouse Mab (3G6) 55 kDa IF, IHC, WB 8 A01080 Anti-α-Tubulin Mouse Mab (3G5) 55 kDa WB, IHC, IF, IP   A01130 Anti-Cyclophilin B Mouse Mab (7B2) 24 kDa WB, IHC, IF  

  If you have any further interests to learn Abbkine loading control antibody category, kindly refer at https://www.abbkine.com/product-category/antibody/loading-control/ , send an E-mail to [email protected], our representative will surely contact you back shortly.

  About Abbkine Scientific Co., Ltd.

Abbkine full portfolio of loading control and epitope tag antibodies

With full portfolio of loading control and epitope tag antibodies, Abbkine launched series of conjugated antibodies, including HRP, Biotin, FITC, Cy3 and AbFluor™ dyes to meet the diversified demands of researchers. Also, the agarose and magnetic beads conjugated tag antibodies specifically for immunoprecipitation are now available.

New Post has been published on https://www.abbkine.com/abbkine-full-portfolio-of-loading-control-and-epitope-tag-antibodies/

Abbkine full portfolio of loading control and epitope tag antibodies

With full portfolio of loading control and epitope tag antibodies, Abbkine launched series of conjugated antibodies, including HRP, Biotin, FITC, Cy3 and AbFluor™ dyes to meet the diversified demands of researchers. Also, the agarose and magnetic beads conjugated tag antibodies specifically for immunoprecipitation are now available.

New Post has been published on https://www.abbkine.com/abbkine-full-portfolio-of-loading-control-and-epitope-tag-antibodies/

Abbkine full portfolio of loading control and epitope tag antibodies

With full portfolio of loading control and epitope tag antibodies, Abbkine launched series of conjugated antibodies, including HRP, Biotin, FITC, Cy3 and AbFluor™ dyes to meet the diversified demands of researchers. Also, the agarose and magnetic beads conjugated tag antibodies specifically for immunoprecipitation are now available.

Interests on https://www.abbkine.com/abbkine-full-portfolio-of-loading-control-and-epitope-tag-antibodies/

Abbkine full portfolio of loading control and epitope tag antibodies

Abbkine announces the launch of Anti-α-Tubulin Monoclonal Antibody (3G5)

Popular scientific research company, Abbike Scientific, has announced the release of its latest antibody named the Anti-α-Tubulin Monoclonal Antibody (3G5). Otherwise known as TUBA1 antibody, the product joins the illustrious list of scientific tools and products from the research giant.

The alpha tubulin antibody is part of the family of microtubules of the eukaryotic cytoskeleton, with the alpha and beta tubulins being the major components of microtubules. The alpha tubulin genes are conserved among several species.

TUBA1A is monoclonal and very similar to the Tuba1 genes in mouse and rat. Studies have further shown that TUBA1A expression is majorly found in morphologically differentiated neurologic cells. As one of the three alpha-tubulin genes in a cluster on chromosome 12q, the mutation in TUBA1A leads to lissencephaly type 3 (LIS3).

The antibody has a recombinant protein immunogen with human, mouse and rat reactivity. The applications of the antibody include IF, IHC, IP, WB and users are advised to determine the best working dilutions after series of experiments. However, the suggested starting dilutions are WB 1:5000-10000, IP 1:200.

The antibody was affinity-purified from mouse ascites by affinity-chromatography using specific immunogen, which is probably one of the unique attributes of the product. The antibody is available in a liquid solution, making it easy for researchers to use, manipulation and loading control.

The makers of the antibody have expressly declared that the product is manufactured for research purpose only and should not be used for human or diagnostic purpose.

About Abbkine Scientific

Abbkine Scientific Co. Ltd is a scientific research company founded by a team of enthusiastic and passionate marketing experts and scientists. Established in 2012 in California, the company moved its headquarters to China due to growing demands from Asia Pacific.