Anti-human IgG mAb (MT145), unconjugated

Anti-human IgG mAb (MT145), unconjugated

3850-1-250

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Content

Monoclonal antibody MT145, unconjugated. Supplied at 0.5 mg/ml in PBS with 0.02% sodium azide.

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Anti-human IgG mAb (MT78), ALP


Product specifications

Intended use

This monoclonal antibody enables specific detection of human IgG in immunoassays such as ELISpot, FluoroSpot, and ELISA.

Serum/Plasma samples

Recommendation

MT145 is recommended as capture mAb in ELISA in combination with detection mAb MT78 (product code 3850-9A) for determination of human total IgG. The antibody is also suitable as capture mAb in ELISpot and FluoroSpot for enumeration of B cells secreting IgG subclasses.

Product details

ProductAnti-human IgG mAb (MT145), unconjugated
ApplicationELISpot, FluoroSpot, ELISA
AnalyteIgG
AntibodyMT145
ConjugateUnconjugated
ClonalityMonoclonal
ImmunogenNatvie human IgG
HostMouse
IsotypeIgG2b
ReactivityHuman, Non-human primates
Specificity

Fc part of human IgG

Cross-reactivity

Cross-reacts with IgG from rhesus macaque and cynomolgus macaques. For information on reactivity with other non-human primates (NHP), please check the NHP cross-reactivity guide.

Purification

Purified from in vitro cultures by protein G affinity chromatography.

Concentration0.5 mg/ml
Supplied in

PBS with 0.02% sodium azide. Sterile-filtered (0.2 µm).

Contents

Monoclonal antibody MT145, unconjugated. Supplied at 0.5 mg/ml in PBS with 0.02% sodium azide.

Shipping and Storage

Shipping

Shipped at ambient temperature.

Storage

Store product at 4-8°C or frozen at -20°C or below. Avoid repeated freezing/thawing.

Shelf lifeAt least 18 months from date of receipt.
Datasheet/Protocol
SDS

Learn how to combine Mabtech’s antibodies for human IgG ELISA and ELISpot. Adjust the setup to detect either total IgG or antigen-specific IgG. Different antibody combinations of monoclonal antibodies have been selected for the detection of the human IgG subclasses in ELISA and ELISpot. There is no cross-reactivity to human IgA1, IgA2, IgM, and IgE.

 

Application guide: ELISA

The human IgG ELISA can be adapted to detect antigen-specific IgG or the total IgG in serum or plasma samples.

Antigen-specific IgG

Capture: Antigen
Detection mAbs: MT78/145, biotin or MT78, ALP

elisa igg 1

Total IgG

Capture mAb: MT145, unconjugated
Detection mAb: MT78, ALP

elisa igg 2

 

Application guide: ELISpot

The human IgG ELISpot and FluoroSpot can be adapted to detect antigen-specific IgG-secreting cells or the total IgG-secreting cells in cell samples like PBMCs.

Antigen-specific IgG

Capture:
Antigen
Detection mAbs:
MT78/145, biotin

elispot human igg 1

 

Capture mAbs:
MT91/145, unconjugated
Detection:
Antigen-biotin

elispot human igg 2

Total IgG

Capture mAbs:
MT91/145, unconjugated
Detection mAbs:
MT78/145, biotin

elispot human igg 3

 

 

Get inspired!

Make sure to check out how researchers have used Mabtech's human IgG ELISA, ELISpot, and FluoroSpot kits in everything from vaccine development to SARS-CoV-2 research. You can find it all in our Publication database!

Find out which analyte combinations we have evaluated in T cell FluoroSpot and which combinations are affected by capture effects or not. 

What is a capture effect?

When capture antibodies with different specificities are coated together, the capture of one cytokine may affect the secretion of other cytokines. This is usually more pronounced when studying T cell responses with polyclonal stimuli compared to antigen-specific responses. Capture effects are seldom a problem and can often be counteracted by the addition of an anti-CD28 antibody.

Depiction of capture effects observed in FluoroSpot

(1) IL-2 secreted by the activated T cell is captured by coated anti-IL-2 capture antibodies. (2) As a result, IL-2-stimulation of the T cell itself (autocrine stimulation) as well as nearby T cells (paracrine stimulation) is impaired, ultimately leading to (3) decreased IFN-γ secretion.

Co-stimulation with anti-CD28

Anti-CD28 mAb provides a co-stimulatory signal to antigen-specific responses by binding to CD28 on T cells. The addition of an anti-CD28 mAb to the cell culture enhances antigen-specific responses and can counteract capture effects. For example, the presence of IL-2 capture antibodies may result in reduced activation of T cells, as capturing of IL-2 decreases the amount of available IL-2 and thereby dampens the IL-2-mediated stimulation of T cells. The addition of anti-CD28 mAb restores IFN-γ responses (depicted in the below images). Further optimization may be necessary, depending on the cells and stimuli used. Too high concentrations of the anti-CD28 mAb may result in an elevation of non-specific cytokine secretion.

Overcoming capture effects using anti-CD28 anitbodies in FluoroSpot

(1) An anti-CD28 antibody can be added to provide a co-stimulatory signal that can restore (2) e.g. IFN-γ responses.

How to investigate capture effects

Our FluoroSpot Plus kits are evaluated for capture effects, and in studies of T cell responses we recommend co-stimulation with anti-CD28. With FluoroSpot Flex, it is possible to combine and build your own kit. For guidance look at our analyte combination table (above). Capture effects can be investigated by quantifying spot numbers in wells coated with single capture antibodies and compared to wells coated with a mixture of the capture antibodies. The compensatory effect of the anti-CD28 mAb may be assessed by comparing cells cultured with and without the anti-CD28 mAb.

The systems reactive with NHPs are either based on cross-reactive human kits or specifically developed monkey kits (NHP).

Cross-reactivity verification tests have been performed in ELISpot and/or ELISA by Mabtech and/or by others. Evaluations of assays with less solid evidence of cross-reactivity are shown as symbols within parentheses.

 Rhesus MacaqueCynomolgus MacaquePig-tailed MacaqueBaboon*Sooty MangabeyAfrican Green MonkeyNight Monkey*Common MarmosetSquirrel Monkey
ASSAYOld worldNew world
ApoA1      
ApoB       
ApoE (NHP)       
ApoE       
ApoH       
CCL2 (MCP-1)       
CCL4 (MIP-1β) ( ) ( )( )( )( )
CCL22 (MDC) ( ) ( )( )( )( )
CD25       
GM-CSF( )( )   
Granzyme A        
Granzyme B (NHP)       
Granzyme B     
IFN-α2        
IFN-α pan   ( )   
IFN-γ (NHP)
IFN-γ( )
IgA (NHP)       
IgA       
IgG       
IgM       
INS     
IL-1α        
IL-1β        
IL-2 (NHP)
IL-3       
IL-4( )( )( )( )
IL-5( )( )( ) 
IL-6( )( ) ( ) 
IL-8 (NHP)        
IL-8 (ELISA)        
IL-8 (ELISpot)        
IL-10 (NHP)      
IL-10       
IL-12/-23 (p40)      
IL-12 (p70)        
IL-13 (NHP)( )( )( )  
IL-13        
IL-17A (NHP)( )( )   
IL-17A     
IL-17F        
IL-17A/F        
IL-21      
IL-22        
IL-23     
IL-27        
IL-31        
IP-10       
Perforin( )   
TGF-β1 (latent)       
TNF-α (NHP)( )  ( )( )( )( )
TNF-α       
          
SINGLE mAbs         
Cell stimulation         
CD3, mAb CD3-1       
CD3, mAb CD3-2       
CD28, mAb CD28-A       
Flow cytometry          
IFN-γ mAb 1-D1K( )( )( )( )( )( )
IL-2 mAb MT8G10( )( )( )( ) ( )( )( )
IL-4 mAb IL4-3( )       
IL-17A mAb MT504        
Perforin mAb Pf-344     
TNF-α mAb MT15B15       
Ig reagents         
IgG1 mAb MTG1218       
IgG1 mAb MT1939       
IgG2 mAb H6200       
IgG2 mAb MTG211E       
IgG3 mAb MTG34       
IgG4 mAb MTG42       
Western blot         
IFN-γ mAb MT111W( )( )( )( )( )( )( )( )( )
TNF-α mAb MT15B15( )( )( )  ( )( )( )( )

Symbol keys:


Cross-reactivity

Verified by Mabtech and/or by others. Tests performed in ELISpot and/or ELISA.

  • good
  • poor
  • no

Indications of Cross-reactivity

Based on reports from others or by analysis using recombinant proteins.

  • ( ) good
  • ( ) poor
  • ( ) no

Monkey kits

Specifically developed monkey kits are marked with (NHP).

Blank boxes

Represent cross-reactivities not evaluated.

* Comprises several species. Cross-reactivity may have to be verified on a species basis.

Tutorial, Published December 28, 2023

Incubation strategies for success in ELISpot and FluoroSpot
We've made a handy table summarizing incubation times for different analytes to make your research planning for the ELISpot and FluoroSpot assay a little easier.

Highlighted research, Published May 30, 2020

B cell FluoroSpot for sensitive ASC detection
Mabtech IRIS and IgG/IgA/IgM FluoroSpot were used to study antibody-secreting plasma cells (ASCs) specific for the nucleocapsid (N) protein. FluoroSpot was shown to be more sensitive than e.g., ELISA.

Highlighted research, Published October 15, 2019

Detection of tumor infiltrating B cells
Tumor infiltrating B cells are able to actively secrete immunoglobulins as shown by performing an IgA, IgG and IgM FluoroSpot. The FluoroSpot technology enabled investigation of the functionality of cells directly isolated from the tumor.

Highlighted research, Published August 18, 2018

A B-cell FluoroSpot assay to assess the risk of rejection
Reversed B-cell FluoroSpot enables the simultaneous study of multiple antigen specificities in the one well. THe assay has been used for Malaria and Dengue virus research, and now for the first time in the context of organ transplantation.
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IgG

Analyte description

Immunoglobulin G (IgG) is the most abundant Ig isotype in serum, making up approximately 80% of all serum immunoglobulins. In humans, there are four subclasses of immunoglobulin G, with the highest serum concentrations of IgG1 followed by IgG2, IgG3, and IgG4. In mice, the IgG subclasses are defined as IgG1, IgG2a/c, IgG2b, and IgG3. The IgG molecule consists of two heavy and two light chains (κ or λ), resulting in a molecule with two arms for antigen binding. High levels of IgG antibodies are induced following the initial IgM response in a typical immune response to antigens.

Alternative namesImmunoglobulin G, IgG, IgG3
Cell typeB cell
Gene ID3500, 3501, 3502
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ELISA Flex: Human IgG (ALP)