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Suitable for ELISA, IHC-P, WB. Ideal for western blot. Cited in 81 publications.


Images

Key facts

Host species

Mouse

Target species

Rabbit

Target isotype

IgG

Target specificity

Light chain

Conjugation

HRP

Applications

ELISA, IHC-P, WB

Clonality

Monoclonal

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Reactivity data

Application

ELISA

Reactivity

Reacts

Dilution info

1/4000.00000 - 1/8000.00000

Notes

-

Application

IHC-P

Reactivity

Reacts

Dilution info

1/8000

Notes

-

Application

WB

Reactivity

Reacts

Dilution info

-

Notes

Note: This antibody is not recommended for sensitive or quantitative detection of the reduced and denatured Rabbit Light Chains on Western Blots.

Alternative names

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Suitable for ELISA, IHC-P, WB. Ideal for western blot. Cited in 81 publications.

Alternative names

Key facts

Description

Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP)

Host species

Mouse

Target species

Rabbit

Target isotype

IgG

Target specificity

Light chain

Conjugation

HRP

Applications

ELISA, IHC-P, WB

Clonality

Monoclonal

Clone number

SB62a

Pre-adsorbed

No

Specificity

ab99697 reacts with the light chain of native Rabbit IgG primary antibodies and does not bind to the reduced and denatured Rabbit IgG heavy chain band (50kD).

Isotype

IgG2b

Light chain type

lambda

Concentration
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Properties

Form

Liquid

Storage buffer

pH: 7.4
Constituents: 50% Glycerol (glycerin, glycerine), 50% PBS

Purification technique

Affinity purification Protein A

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

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7 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

    IHC image of beta actin staining in human colon formalin fixed paraffin embedded tissue section*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab8227, 3μg/ml overnight at +4°C. An HRP-conjugated secondary (ab99697, 1/8000 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.

    The inset negative control image is secondary-only at 1/4000 dilution.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697), expandable thumbnail

    Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

    LAP2 alpha was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to LAP2 alpha (ab5162)and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Hela whole cell extract diluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab5162. Secodary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Bands: 83kDa: LAP2 alpha.

    Secondary

    All lanes: Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (AB99697)

  • Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697), expandable thumbnail

    Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

    Western blot showing that Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) specifically binds to light-chains


    Left blot: Coomassie stain showing the rabbit heavy (~50kDa) and light-chains (~25 kDa)

    Lane 1: 10 µg rabbit monoclonal was loaded

    Lane 2: 5 µg rabbit monoclonal was loaded

    Lane 3: 2 µg rabbit monoclonal was loaded

    Right blot: WB using Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) as secondary antibody at 1/10,000

    Lane 1: 1 µg rabbit monoclonal was loaded

    Lane 2: 0.5 µg rabbit monoclonal was loaded

    Lane 3: 0.25 µg rabbit monoclonal was loaded

    Secondary

    All lanes: Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (AB99697)

  • Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697), expandable thumbnail

    Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

    Catalase was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Catalase (ab1877) and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Hela whole cell extract diluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab1877. Secodary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
    Bands: 60kDa: Catalase

    Secondary

    All lanes: Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (AB99697)

  • Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697), expandable thumbnail

    Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)


    All lanes : Anti-beta Actin antibody - Loading Control (ab8227) at 1 μg/ml

    Lanes 1-6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate (ab27252)
    Lysates/proteins at 10 μg per lane.

    Secondary
    Lanes 1 - 2 : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) at 1/2000 dilution
    Lanes 3 - 4 : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) at 1/10000 dilution
    Lanes 5 - 6 : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.
    Secondary

    All lanes: Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (AB99697)

  • Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697), expandable thumbnail

    Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

    Rad21 was immunoprecipitated using 0.5mg Hela whole cell extract, 5ug of Rabbit polyclonal to Rad21and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Hela whole cell extractdiluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab42478. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) .. Band: 100kDa: Rad21; Non specific - 50kDa: We are unsure as to the identity of this extra band.

    Although the predicted band size is 75kDa based on Swiss-prot data, a band of 120kDa has been previously observed. Mol Cell Biol. 2002 Dec;22(23):8267-77 PMID: 12417729. Th

    Secondary

    All lanes: Immunoprecipitation - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (AB99697)

  • Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697), expandable thumbnail

    Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (ab99697)

    Lane 1: Rabbit IgG loaded at 1ug/well

    ab99697 diluted to 1:2000

    Secondary

    All lanes: Western blot - Mouse monoclonal [SB62a] Anti-Rabbit IgG light chain (HRP) (AB99697)

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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