Anti-Kappa light chain antibody [RM1057] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Kappa light chain antibody [RM1057] - BSA and Azide free (AB313896)

This data was developed using ab313895, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Raji (human Burkitt's lymphoma B lymphocyte) cells labelling Kappa light chain with ab313895 at 1/500 (0.908 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing cytoplasmic staining in Raji cell line. Negative control : MCF7. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kappa light chain antibody [RM1057] - BSA and Azide free (AB313896)

This data was developed using ab313895, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Kappa light chain with ab313895 at 1/5000 (0.227 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on stromal cells of human breast cancer.The section was incubated with ab313895 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Kappa light chain antibody [RM1057] - BSA and Azide free (AB313896)

This data was developed using ab313895, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Kappa light chain with ab313895 at 1/5000 (0.227 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human tonsil.The section was incubated with ab313895 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

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Immunoprecipitation - Anti-Kappa light chain antibody [RM1057] - BSA and Azide free (AB313896)

This data was developed using ab313895, the same antibody clone in a different buffer formulation. Kappa light chain was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate with ab313895 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313895 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate Lane 2 : ab313895 IP in Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313895 in Raji whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 180 seconds

All lanes:

Immunoprecipitation - Anti-Kappa light chain antibody [RM1057] (<a href='/en-us/products/primary-antibodies/kappa-light-chain-antibody-rm1057-ab313895'>ab313895</a>) at 1/30 dilution

All lanes:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• WB

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Western blot - Anti-Kappa light chain antibody [RM1057] - BSA and Azide free (AB313896)

This data was developed using ab313895, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : cerebellum. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 48 seconds

All lanes:

Western blot - Anti-Kappa light chain antibody [RM1057] (<a href='/en-us/products/primary-antibodies/kappa-light-chain-antibody-rm1057-ab313895'>ab313895</a>) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human kidney tissue lysate at 20 µg

Lane 4:

Human spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Observed band size: 25 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-Kappa light chain antibody [RM1057] - BSA and Azide free (AB313896)

This data was developed using ab313895, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : MCF7 In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 3 minutes

All lanes:

Western blot - Anti-Kappa light chain antibody [RM1057] (<a href='/en-us/products/primary-antibodies/kappa-light-chain-antibody-rm1057-ab313895'>ab313895</a>) at 1/1000 dilution

Lane 1:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Observed band size: 25 kDa

false

Exposure time: 3s

• I-ELISA

Supplier Data

Indirect ELISA - Anti-Kappa light chain antibody [RM1057] - BSA and Azide free (AB313896)

This data was developed using ab313895, the same antibody clone in a different buffer formulation. Indirect ELISA analysis of ab313895 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution. Antigen : Human Kappa Light Chains, Human Lamda Light Chains, Human IgA, Human IgM, Human IgG, Rat IgG, Mouse IgG. Antigen concentration : 1000 ng/ml