Revision 4

#82042Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IHC-P

REACTIVITY:

M

SENSITIVITY:

Endogenous

MW (kDa):

80

Source/Isotype:

Rabbit IgG

UniProt ID:

#P01872

Entrez-Gene Id:

16019

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunohistochemistry (Paraffin) 1:100 - 1:400

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #49314.

Specificity / Sensitivity

IgM (E8M1B) XP® Rabbit mAb recognizes endogenous levels of total IgM heavy chain protein. This antibody reacts with secreted and membrane bound isoforms of IgM heavy chain protein.

Species Reactivity:

Mouse

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of the constant region of mouse IgM heavy chain protein.

Background

Immunoglobulin M (IgM) is one of the five major human immunoglobulin isotypes and is the first antibody to be secreted by plasmablasts in a humoral immune response after exposure to antigen. Structurally, IgM is the largest immunoglobulin and predominantly exists in pentameric form when secreted (1). Alternative splicing of the IgM heavy chain mRNA can generate an alternative form of the antibody, which facilitates its insertion into the plasma membrane of B cells to function in antigen recognition. IgM is the first Ig isotype to appear on the surface of developing B cells and is a major component of the B cell antigen receptor (BCR) signaling complex, which drives B cell survival and proliferation upon antigen-induced ligation (2,3). Research studies have shown that defects in the assembly of the BCR account for the low levels of surface IgM expression in B-chronic lymphocytic leukemia (4,5).

  1. Fazel, S. et al. (1997) Int Immunol 9, 1149-58.
  2. Tseng, J. et al. (1994) Immunol Res 13, 299-310.
  3. DeFranco, A.L. et al. (1995) Ann N Y Acad Sci 766, 195-201.
  4. Vuillier, F. et al. (2005) Blood 105, 2933-40.
  5. Payelle-Brogard, B. et al. (2002) Br J Haematol 118, 976-85.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IHC-P: Immunohistochemistry (Paraffin)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 4
#82042

IgM (E8M1B) XP® Rabbit mAb

Western Blotting Image 1: IgM (E8M1B) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from WEHI-231 cells, wild-type BALB/c mouse splenocytes (wt/wt), Jh knockout (KO) BALB/c mouse splenocytes (-/-, model #1147-F), and C2C12 cells using IgM (E8M1B) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Jh (BALB/c) mouse from Taconic Biosciences, Inc. The Jh mouse carries a homozygous deletion of the J segments of the Ig heavy chain locus, which results in defective heavy chain recombination and lack of IgM due to a deficiency in mature B cells. Negative expression of IgM protein in C2C12 cells is consistent with the predicted expression pattern.
Immunohistochemistry Image 1: IgM (E8M1B) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded A20 syngeneic tumor using IgM (E8M1B) XP® Rabbit mAb.
Immunohistochemistry Image 2: IgM (E8M1B) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mammary tumor using IgM (E8M1B) XP® Rabbit mAb.
Immunohistochemistry Image 3: IgM (E8M1B) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded lung from BALB/c (wt/wt, left) and Jh knockout (-/-, model #1147-F, right) mice using IgM (E8M1B) XP® Rabbit mAb. Jh (BALB/c) mouse from Taconic Biosciences, Inc.
Immunohistochemistry Image 4: IgM (E8M1B) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded spleen from BALB/c (wt/wt, left) and Jh knockout (-/-, model #1147-F, right) mice using IgM (E8M1B) XP® Rabbit mAb. Jh (BALB/c) mouse from Taconic Biosciences, Inc.
Immunohistochemistry Image 5: IgM (E8M1B) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded spleen (left), lung (middle), or thymus (right) from BALB/c (wt/wt, top) and Jh knockout (-/-, model #1147-F, bottom) mice using IgM (E8M1B) XP® Rabbit mAb. Jh (BALB/c) mouse from Taconic Biosciences, Inc.
Immunohistochemistry Image 6: IgM (E8M1B) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse kidney using IgM (E8M1B) XP® Rabbit mAb.
Immunohistochemistry Image 7: IgM (E8M1B) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse spleen using IgM (E8M1B) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 8: IgM (E8M1B) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded WEHI-231 cell pellet (left, positive) or C2C12 cell pellet (right, negative) using IgM (E8M1B) XP® Rabbit mAb.