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 Anti-IgG, Light Chain Specific for Western Blotting:

Anti-IgG, Light Chain Specific
for Protein Detection on Western Blots after Immunoprecipitation

When labeled secondary antibodies specific for both heavy and light chains of IgG, e.g. anti-IgG (H+L), are used to detect protein bands on Western blots following immunoprecipitation (IP), two heavy bands appear (Figure A) corresponding to the heavy (50 kDa) and light chains (25 kDa) of the precipitated primary antibody. These bands usually obscure detection of any protein of interest with a molecular weight near 50 kDa or 25 kDa. However, when labeled anti-IgG, Light Chain Specific antibodies are used for detection, they bind only to the light chain band on the blot (Figure B) and to light chains on the native primary antibodies used for detection. Therefore, a 50 kDa protein may be detected on blots without interference from the precipitated IgG in the same band by incubation with its unlabeled primary antibody followed by labeled anti-IgG, Light Chain Specific. Similarly, a 25 kDa protein may be detected after IP without interference by using its unlabeled primary antibody followed by labeled anti-IgG, Fc fragment specific.

       

Figures A+B. Heavy (50kDa) and light (25kDa) chains of reduced and SDS-denatured mouse IgG were separated by SDS-PAGE and detected on Western blots using HRP-goat anti-mouse IgG (H+L) (A) and HRP-goat anti-mouse IgG, LC specific (B). Both heavy and light chain bands were detected with anti-IgG (H+L) (A). However, no heavy chain band was detected when anti-IgG, LC specific antibodies were used (B) even on lanes heavily overloaded with IgG.

Anti-IgG, Light Chain Specific antibodies also have been thoroughly adsorbed to minimize cross-reactivity with immunoglobulins from other species which may be present on blots.

Although the antibodies react strongly with native IgG light chains, some do not react as strongly with reduced and denatured light chains. Therefore, they are not recommended for sensitive detection and quantitation of reduced and denatured light chains on Western blots.

 
 

 

Antibody Description Unconjugated Cyanine
Cy2
A=492, E=510

DyLight
488

A=493, E=518

Alexa Fluor®
488
A=493, E=519

DyLight
549

A=555, E=568
Cyanine
Cy3

A=550, E=570

Alexa Fluor®
594

A=591, E=614

DyLight
594

A=591, E=616

Alexa Fluor®
647
A=651, E=667

DyLight
649

A=652, E=670
NEW
Alexa Fluor®
680
A=684, E=702
NEW
Alexa Fluor®
790
A=792, E=803
Biotin-SP
(long spacer)
Horseradish
Peroxidase
Alkaline
Phosphatase
  IgG Fraction Monoclonal Mouse Anti-Goat IgG,
  Light Chain
Specific
  (min X Hrs, Hu, Ms, Rb, Rat Ig)
$112 / 1.0 mg
205-002-176
  $184 / 0.5 mg
205-482-176
$184 / 0.5 mg
205-542-176
  $160 / 0.5 mg
205-162-176
$184 / 0.5 mg
205-582-176
  $ 184 / 0.5 mg
205-602-176
$184 / 0.5 mg
205-492-176
$167 / 0.5 mg
205-622-176
$167 / 0.3 mg
205-652-176
$143 / 0.5 ml
205-062-176
$143 / 0.5 ml
205-032-176
$158 / 0.5 ml
205-052-176
  AffiniPure Goat Anti-Mouse IgG Light Chain * Specific
  (min X Bov, Gt, Hrs, Hu, Rb, Rat, Shp Ig)
112 / 1.0
115-005-174

 

 

184 / 0.5
115-545-174

$ 184 / 0.5 mg
115-505-174

160 / 0.5
115-165-174
184 / 0.5
115-585-174

$ 184 / 0.5 mg
115-515-174

184 / 0.5
115-605-174

 

167 / 0.3
115-625-174
167 / 0.3
115-655-174
143 / 0.5
115-065-174
143 / 0.5
115-035-174
158 / 0.5
115-055-174
  IgG Fraction Monoclonal Mouse Anti-Rabbit IgG,
  Light Chain
Specific
  (min X Bov, Gt, Arm Hms, Hrs, Hu, Ms, Rat, Shp Ig)
112 / 1.0
211-002-171
$ 160 / 0.5 mg
211-222-171
184 / 0.5
211-482-171
184 / 0.5
211-542-171
184 / 0.5
211-502-171
160 / 0.5
211-162-171
184 / 0.5
211-582-171
184 / 0.5
211-512-171
184 / 0.5
211-602-171
184 / 0.5
211-492-171
167 / 0.5
211-622-171
167 / 0.3
211-652-171
143 / 0.5
211-062-171
143 / 0.5
211-032-171
158 / 0.5
211-052-171
  AffiniPure Goat Anti-Rat IgG, Light Chain*Specific
  (min X Bov, Gt, Hrs, Hu, Ms, Rb, Shp Ig)
112 / 1.0
112-005-175
160 / 0.5
112-225-175
184 / 0.5
112-485-175
184 / 0.5
112-545-175
184 / 0.5
112-505-175
160 / 0.5
112-165-175
184 / 0.5
112-585-175
184 / 0.5
112-515-175
184 / 0.5
112-605-175
184 / 0.5
112-495-175
167 / 0.3
112-625-175
167 / 0.3
112-655-175
143 / 0.5
112-065-175
143 / 0.5
112-035-175
158 / 0.5
112-055-175
  IgG Fraction Monoclonal Mouse Anti-Sheep IgG,
  Light Chain
Specific
  (min X Bov, Hrs, Hu, Ms, Rb, Rat Ig)
112 / 1.0
213-002-177
    184 / 0.5
213-542-177
  160 / 0.5
213-162-177
184 / 0.5
213-582-177
  184 / 0.5
213-602-177
184 / 0.5
213-492-177
167 / 0.3
213-622-177
167 / 0.3
213-652-177
143 / 0.5
213-062-177
143 / 0.5
213-032-177
158 / 0.5
213-052-177

 Warning: BSA and dry milk may contain IgG which will react with this antibody.  Use of BSA and/or dry milk to block or dilute this antibody may increase background and/or reduce secondary antibody titer.
*
 
This antibody reacts primarily with kappa light chains. It is not suitable for detection of primary antibodies with lambda light chains.

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